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Research ArticleEffects of Methanol Extract of Breadfruit (Artocarpus altilis)on Atherogenic Indices and Redox Status of Cellular System ofHypercholesterolemic Male Rats
Oluwatosin Adekunle Adaramoye and Olubukola Oyebimpe Akanni
Drug Metabolism and Toxicology Research Laboratories Department of Biochemistry College of MedicineUniversity of Ibadan Ibadan 20005 Nigeria
Correspondence should be addressed to Oluwatosin Adekunle Adaramoye aoadaramoyeyahoocom
Received 23 November 2013 Revised 17 December 2013 Accepted 18 December 2013 Published 30 January 2014
Academic Editor Neal Davies
Copyright copy 2014 O A Adaramoye and O O Akanni This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited
We investigated the effects of methanol extract of Artocarpus altilis (AA) on atherogenic indices and redox status of cellular systemof rats fed with dietary cholesterol while Questran (QUE) served as standard Biochemical indices such as total cholesterol (TC)triglycerides (TG) low- and high-density lipoproteins-cholesterol (LDL-C and HDL-C) aspartate and alanine aminotransferases(AST and ALT) lactate dehydrogenase (LDH) reduced glutathione glutathione-s-transferase glutathione peroxidase (GPx)catalase (CAT) superoxide dismutase (SOD) and lipid peroxidation (LPO) were assessed Hypercholesterolemic (HC) rats hadsignificantly increased relative weight of liver and heart Dietary cholesterol caused a significant increase (119875 lt 005) in the levelsof serum hepatic and cardiac TC by 110 70 and 85 LDL-C by 79 82 and 176 and TG by 68 96 and 62respectively Treatment with AA significantly reduced the relative weight of the organs and lipid parameters There were beneficialincreases in serum and cardiac HDL-C levels in HC rats treated with AA In HC rats serum LDH ALT and AST activities andlevels of LPO were increased whereas hepatic and cardiac SOD CAT and GPx were reduced All biochemical and histologicalalterations were ameliorated upon treatment with AA Extract of AA had protective effects against dietary cholesterol-inducedhypercholesterolemia
1 Introduction
Recent studies have demonstrated that increased formationof free radicalsreactive oxygen species (ROS) contributesto cardiovascular disease (CVD) progression [1] Generationof large amounts of ROS can overwhelm the intracellularantioxidant defense causing lipid peroxidation proteinmod-ification and DNA breaks [2] It is known that ROS-induceddepletion of antioxidants is a key factor for the initiation ofatherosclerosis and the development of CVD [3] Hyperc-holesterolemia characterized by the presence of high levels ofcholesterol in the blood [4] is a form of hyperlipidemia andhyperlipoproteinemia Hyperlipidemia has also been foundto induce oxidative stress in various organs of the body[5] Although several factors such as life style a diet richin cholesterol and age have been reported to cause heart
failure [6 7] high levels of cholesterol particularly LDL-cholesterol are mainly responsible for hypercholesterolemia[8] Drugs that lower cholesterol such as fibrates and bileacid sequestrants were used for several decades but the highprevalence of adverse effects led to the introduction of statins(HMG-CoA inhibitors) [9] Although the adverse effect ofstatins is relatively low one rare effect called rhabdomyolysiscan be very serious with statins [9] In view of these adverseeffects the quest for natural products with hypolipidemicpotential and minimal side effect is warranted
Breadfruit (Artocarpus altilis) is a flowering tree in themulberry family The fruit can be eaten once cooked or canbe further processed into a variety of other foods It is anexcellent source of fiber calcium copper iron magnesiumpotassium thiamine niacin carbohydrates and vitaminsand very low in fat [10] In herbal homes leaves of this plant
Hindawi Publishing CorporationAdvances in Pharmacological SciencesVolume 2014 Article ID 605425 11 pageshttpdxdoiorg1011552014605425
2 Advances in Pharmacological Sciences
are used for the treatment of liver disorders hypertensionand diabetes [11 12] In vitro studies by Nwokocha et al[13] supported the folkloric use of this plant However littleinformation is available with respect to in vivo studies on itsethnomedicinal usesThis study was designed to evaluate theeffects of Artocarpus altilis on atherogenic indices and redoxstatus of cellular system of hypercholesterolemic rats
2 Materials and Methods
21 Chemicals Questran (Bristol-Myers Squibb HounslowUK) was purchased from a local chemist in IbadanNigeria Dietary cholesterol and thiobarbituric acid (TBA)were procured from Aldrich Chemical Co (MilwaukeeWI USA) Glutathione hydrogen peroxide 551015840-dithio-bis-2-nitrobenzoic acid (DNTB) and epinephrine were pur-chased from Sigma Chemical Co Saint Louis MO USATrichloroacetic acid (TCA) and thiobarbituric acid (TBA)were purchased from British Drug House (BDH) ChemicalLtd Poole UK Other reagents were of analytical grade andthe purest quality available
22 Collection and Extraction of Artocarpus altilis The stembark of Artocarpus altilis was collected in Ibadan (Oyo State)and authenticated at the BotanicalGarden of theUniversity ofIbadan The stem bark of Artocarpus altilis was air-dried andcrushed into fine powder The powdered part was extractedwith n-hexane and methanol using soxhlet extractor andthe extract was concentrated in vacuum at 40∘C with rotaryevaporator and water bath to dryness The yield of theextraction was 57
23 Determination of Total Phenolic Contents The totalphenolic content of the extract was determined using themethod of Singleton et al [14] with slight modificationsFolin-C reagent (1mL) was added to 1mL of extract orstandard After 3 minutes 1mL of 15 Na
2CO3was added
and the solution was made up to 5mL with distilled waterThe reaction mixture was kept in the dark for 90 minuteswith intermittent shaking or placed in a water bath at 40∘Cfor 20 minutes The absorbance was measured by a BeckmanDU (70) Spectrophotometer at 760 nm All experimentswere done in triplicate A standard curve was plotted withcatechin and the phenolic content expressed as CE (catechinequivalent) per mg dry weight of the extract
231 DPPHmdashRadical Scavenging Activity The radical scav-enging activity of the extract was measured as described byMensor et al [15] The stable 22-diphenyl-1-picrylhydrazyl(DPPH) radical was used for the determination of freeradical scavenging activities of the extracts A portion (1mL)of each of the different concentrations (10ndash1000 120583gmL) ofthe extracts or standard (catechin) was added to 1mL of1mM DPPH in methanol The mixtures were vortexed andincubated in a dark chamber for 30 minutes after which theabsorbance was measured at 517 nm against a DPPH controlcontaining only 1mL of methanol in place of the extract All
calculations were carried out in triplicates The inhibition ofDPPH was calculated as a percentage using the expression
119868 =119860control minus 119860 sample
119860controltimes 100 (1)
where 119868 is the percentage inhibition of the DPPH radical119860control is the absorbance of the control and 119860 sample is theabsorbance of the test compound
24 Animals Inbred male Wistar rats weighing between150 and 180 g were purchased from the animal house ofthe Department of Veterinary Physiology Biochemistry andPharmacology University of Ibadan Nigeria Animals werekept in ventilated cages at room temperature (28ndash30∘C) andmaintained on normal laboratory chow (Ladokun FeedsIbadan Nigeria) and water ad libitum Rats handling andtreatments conform to guidelines of the National Institute ofHealth (NIH publication 85-23 1985) for laboratory animalcare and use The study was approved by the Faculty ofBasic Medical Sciences University of Ibadan Animal EthicsCommittee
25 Study Design Thirty-five male rats were randomlydivided into seven groups of five rats each The first group(control) received drug vehicle (corn oil) the second group(HC) received dietary cholesterol at 30mg03mL [16]the third group (HC + AA1) received dietary cholesteroland Artocarpus altilis (100mgkg) the fourth group (HC+ AA2) received dietary cholesterol and Artocarpus altilis(200mgkg) the fifth group (HC + QUE) received dietarycholesterol and Questran (026 gkg) [16] the sixth group(QUE) received questran alone and the seventh group (AA)receivedArtocarpus altilis at a dose of 200mgkg bodyweight
26 Preparation of Tissues Rats were fasted overnight andsacrificed 24 hours after the last dose of drugs Liver andheart were quickly removed and washed in ice-cold 115KCl solution dried and weighed A section of liver andaorta samples were fixed in 10 formalin for histologicalexamination The remaining parts of liver and heart werehomogenized in 4 volumes of 50mM phosphate buffer pH74 and centrifuged at 10000 g for 15 minutes to obtain post-mitochondrial supernatant fraction (PMF) All procedureswere carried out at temperature of 0ndash4∘C
261 Preparation of Serum Blood was collected from theheart of the animals into plain centrifuge tubes and wasallowed to stand for 1 hour Serum was prepared by cen-trifugation at 3000 g for 15 minutes in a Beckman benchcentrifugeThe clear supernatant was used for the estimationof serum lipid profile and enzymes
27 Biochemical Assays Protein contents of the samples wereassayed by the method of Lowry et al [17] using bovineserum albumin as standard The activities of alanine andaspartate aminotransferases (ALT and AST) were assayed bythe combinedmethods ofMohun andCook [18] andReitman
Advances in Pharmacological Sciences 3
Table 1 Changes in the body weight and relative weight of organs of hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
Treatment Body weight (g) Weight of organs (g) Relative weight of organsInitial Final Liver Kidney Heart Liver Kidney Heart
Control 15100 plusmn 494 19400 plusmn 2302 538 plusmn 049 103 plusmn 011 053 plusmn 009 277 plusmn 075 053 plusmn 005 027 plusmn 004
HC 16000 plusmn 310 19800 plusmn 2903 689 plusmn 089 109 plusmn 014 069 plusmn 006 348 plusmn 023lowast055 plusmn 003 039 plusmn 002
lowast
HC + AA1 15900 plusmn 548 19600 plusmn 3715 512 plusmn 122 109 plusmn 020 055 plusmn 008 261 plusmn 013lowastlowast 056 plusmn 004 028 plusmn 003lowastlowast
HC + AA2 16700 plusmn 604 18500 plusmn 4500 520 plusmn 124 109 plusmn 020 052 plusmn 009 281 plusmn 015lowastlowast 059 plusmn 006 028 plusmn 004lowastlowast
HC + QUE 17300 plusmn 474 20250 plusmn 500 539 plusmn 105 119 plusmn 007 055 plusmn 001 266 plusmn 043lowastlowast 059 plusmn 005 027 plusmn 001lowastlowast
QUE 18800 plusmn 395 22500 plusmn 2041 506 plusmn 121 117 plusmn 005 051 plusmn 049 224 plusmn 041 052 plusmn 007 023 plusmn 004
AA2 19200 plusmn 834 23500 plusmn 2236 519 plusmn 055 121 plusmn 011 062 plusmn 006 221 plusmn 017 052 plusmn 004 026 plusmn 003
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
and Frankel [19] Serum total cholesterol level was assayedby the method of Richmond [20] The method involvedenzymatic hydrolysis and oxidation of cholesterol with theformation of quinoneimine (an indicator) from hydrogenperoxide and 4-aminoantipyrine in the presence of phenoland peroxideThe serum level of triglyceride was determinedby Jacobs and van Demark [21] and Koditschek and Umbreit[22] this was based on the hydrolysis of triglycerides with theformation of glycerol which is substrate for other enzymeswith the subsequent formation of hydrogen peroxide Thisthen reacts with 4-aminophenazone and 4-chlorophenol inthe presence of peroxidase to give quinoneimine which ismeasured spectrophotometrically at 500 nm
The lipoproteins (measured using the enzymatic col-orimetric method) very low-density lipoprotein (VLDL)and low-density lipoprotein (LDL) were precipitated by theaddition of phosphotungstic acid and magnesium chlorideAfter centrifugation at 3000 g for 10 minutes at 25∘C theclear supernatant containedHDL fraction whichwas assayedfor cholesterol with the Randox diagnostic kit The low-density lipoprotein (LDL) was calculated using the formulaof Friedewald et al [23] Lipid peroxidation level was assayedby the reaction between 2-thiobarbituric acid (TBA) andmalondialdehyde (MDA) an end product of lipid peroxidesas described by Buege and Aust [24]
The activity of lactate dehydrogenase (LDH) was deter-mined by the method of Zimmerman and Weinstein [25]while tissue superoxide dismutase (SOD) activity was mea-sured by the nitro blue tetrazolium (NBT) reduction methodof McCord and Fridovich [26] Catalase (CAT) activity wasassayed spectrophotometrically by measuring the rate ofdecomposition of hydrogen peroxide at 240 nm as describedby Aebi [27] Reduced glutathione level was measured by themethod of Beutler et al [28] this method is based on thedevelopment of a relatively stable (yellow) colour when 5101584051015840-dithiobis-(2-nitrobenzoic acid) (Ellmanrsquos reagent) is added tosulfhydryl compounds The chromophoric product resultingfrom the reaction of Ellmanrsquos reagent with the reducedglutathione (2-nitro-5-thiobenzoic acid) possesses a molarabsorption at 412 nm which is proportion to the level ofreduced glutathione in the test sample The glutathione per-oxidase (GPx) activity was assessed by themethod of Rotruck
et al [29] while glutathione-S-transferase (GST) activity wasdetermined according to Habig et al [30] the principle isbased on the fact that all of known GST demonstrates arelatively high activity with 1-chloro-24-dinitrobenzene asthe second substrate When this substance is conjugated withreduced glutathione its absorption maximum shifts to alonger wavelength 340 nm and the absorption increase at thiswavelength provides a direct measurement of the enzymaticreaction
271 Determination of Antiatherogenic Cardioprotective andCoronary Risk Indices Cardioprotective index (CPI) wasestimated in terms of HDL-C to LDL-C ratio [31 32] whereasantiatherogenic (AAI) and coronary risk indices (CRI) werecalculated by the following formulae [33 34]
AAI = 100 times [HDL-C][Total cholesterol minusHDL-C ]
CRI = Total cholesterolHDL-cholesterol
(2)
28 Histopathology of Tissues Tissues fixed in 10 formalinwere dehydrated in 95 ethanol and then cleared in xylenebefore embedded in paraffin Microsections (about 4 120583m)were prepared and stained with haematoxylin and eosin(HampE) dye and were examined under a light microscope by ahistopathologist who was ignorant of the treatment groups
29 Statistical Analysis All values were expressed as themean plusmn SD of five animals per group Data were analyzedusing one-way ANOVA followed by the post hoc Duncanmultiple range test for analysis of biochemical data usingSPSS (100) Values were considered statistically significant at119875 lt 005
3 Results
31 Phenolic and Flavonoids Contents and Effects of Arto-carpus altilis on Body Weight and Relative Weight of Organsof Hypercholesterolemic (HC) Rats In Table 1 there weresignificant increases (119875 lt 005) in the relative weight of liver
4 Advances in Pharmacological Sciences
Table 2 The total phenolic contents and scavenging activity ofArtocarpus altilis on 22-diphenyl-1-picrylhydrazyl radical (DPPH)in vitro
Conc Scavenging activity Phenolic content(120583gmL) Catechin AA (120583gCEmg)100 422 plusmn 44 227 plusmn 59 018 plusmn 002
300 476 plusmn 16 504 plusmn 31 039 plusmn 003
500 631 plusmn 51 511 plusmn 23 052 plusmn 007
750 678 plusmn 39 625 plusmn 85 068 plusmn 005
Data are expressed as mean plusmn SD (119899 = 4)AA Artocarpus altilis
and heart of HC rats when compared with the control whiletreatment with AA (100 and 200mgkg) significantly reducedthe relative weight of heart and liver of HC rats to values thatwere statistically similar (119875 gt 005) to the control Similarreductionwas obtained in questran-treatedHC ratsThe totalphenolic contents (TPC) of AA expressed in 120583g catechinequivalent per mg dry weight of the extract increased withincrease in concentration (Table 2) At 750 120583gmL the TPCof AA was 068 plusmn 005 120583gCEmg There were significant(119875 lt 005) and dose-dependent increases in scavengingactivity of AA on DPPH radicals (Table 2) At 100 120583gmLand 750 120583gmL the percentage DPPH radical scavengingactivities of AA were 422 and 678 respectively
32 Effects of Artocarpus altilis on Antioxidant Parametersand Marker Enzymes in Hypercholesterolemic (HC) RatsAdministration of dietary cholesterol significantly increased(119875 lt 005) serum hepatic and cardiac lipid peroxidation(LPO) products measured as thiobarbituric acid reactivesubstances (TBARS) by 265 83 and 80 respectively(Table 3) However treatment with AA completely amelio-rated dietary cholesterol-induced increase in LPO In HCrats the activities of hepatic and cardiac SOD and CATas well as cardiac GPx decreased significantly relative tothe control (Table 4) Specifically hepatic SOD and CATdecreased by 54and 45while cardiac SODCAT andGPxdecreased by 67 59 and 36 respectively Also activitiesof phase II and antioxidant enzyme (GST) in the liver of HCrats were significantly reduced when compared to controls(Figure 5) Administration of AA (200mgkg) reversed theadverse effect of high dietary cholesterol by normalizing theseenzymic antioxidant indices In HC rats serum ALT ASTand LDH were significantly increased by 23- 17- and 24-fold respectively while cardiac LDH activity was decreasedby 30-fold relative to controls (Table 5 and Figures 3 and4) However the observed elevations in the activities of theseserum enzymes in HC rats were reversed following treatmentwith AA and quetsran
33 Effects of Artocarpus altilis on the Lipid Profile of Hyperc-holesterolemic Rats Feeding rats on high dietary cholesterolfor nine consecutive weeks significantly (119875 lt 005) increasedthe serum hepatic and cardiac total cholesterol levels by110 70 and 85 respectively (Table 6 and Figures 1 and2) Furthermore serum hepatic and cardiac triglyceridesincreased by 68 96 and 62 while serum LDL-C
0
200
400
600
800
1000
1200
1400
1600
Con
c (m
gdL
)Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Heart total cholHeart trig
lowast
lowast
lowastlowastlowastlowast
lowastlowast
lowastlowastlowastlowast
Figure 1 Effects of methanol extract of Artocarpus altilis andQuestran on cardiac total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC Hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
increased by 79 respectively in HC rats relative to controlsIn addition HC rats had significantly lower HDL-C valueswhen compared to the control (Table 6) Administration ofAA at 200mgkg attenuated the elevated levels of these lipidindices to near normal in the tissues ofHC ratsTheprotectiveeffect of AA at 200mgkg seems better than the standardhypolipidemic drug (Questran) Furthermore AA increasedserum antiatherogenic index in HC rats while coronary riskindex was decreased (Table 7)
34 Effects of Artocarpus altilis on the Histology of Aorta andLiver Thehistology of liver slide showedmarked portal con-gestion severe periportal cellular infiltration bymononuclearcells and mild diffuse vacuolar degeneration of hepatocytes(Figure 6) while aorta from HC rats revealed large focal areaof myofibril necrosis with severe hemorrhages and fibrousconnective tissue laid down (Figure 7) Treatment withAA (200mgkg) reversed the adverse effect of high dietarycholesterol on the histological architecture of the aorta andliver of the rats The histological results further corroboratedthe biochemical findings indicating the beneficial effects ofAA in hypercholesterolemic rats
4 Discussion
It is generally known that elevation of serum LDL-C and totalcholesterol (TC) can lead to CVD especially atherosclerosis
Advances in Pharmacological Sciences 5
Table 3 Changes in the levels of lipid peroxidation in hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nineweeks
Treatments Liver(120583molMDAmg protein)
Heart(120583molMDAmg protein)
Serum(120583molMDAmg protein)
Control 006 plusmn 001 015 plusmn 002 132 plusmn 024
HC 011 plusmn 002lowast
027 plusmn 003lowast
482 plusmn 068lowast
HC + AA1 008 plusmn 003lowastlowast
013 plusmn 003lowastlowast
263 plusmn 056lowastlowast
HC + AA2 007 plusmn 002lowastlowast
014 plusmn 003lowastlowast
204 plusmn 035lowastlowast
HC + QUE 008 plusmn 002 015 plusmn 002 285 plusmn 077
QUE 005 plusmn 001 016 plusmn 007 182 plusmn 060
AA2 007 plusmn 001 015 plusmn 002 116 plusmn 033
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 4 Changes in the levels of hepatic and cardiac antioxidant parameters in hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
TreatmentLiver Heart
GSH GPx SOD CAT GSH GPx SOD CAT(mgg tissue) (Umg protein) (mgg tissue) (Umg protein)
Control 085 plusmn 015 555 plusmn 083 736 plusmn 101 557 plusmn 108 2106 plusmn 111 14574 plusmn 574 003 plusmn 001 537 plusmn 081
HC 073 plusmn 001 492 plusmn 008 340 plusmn 069lowast
304 plusmn 005lowast1925 plusmn 099 9282 plusmn 287
lowast
001 plusmn 001lowast
219 plusmn 065lowast
HC + AA1 092 plusmn 026 498 plusmn 006 726 plusmn 100lowastlowast 543 plusmn 171lowastlowast 1935 plusmn 085 10843 plusmn 287 001 plusmn 001 299 plusmn 074
HC + AA2 096 plusmn 025 573 plusmn 150 743 plusmn 071lowastlowast 555 plusmn 159lowastlowast 2038 plusmn 111 13747 plusmn 301lowastlowast 003 plusmn 000lowastlowast 403 plusmn 080lowastlowast
HC + QUE 085 plusmn 012 537 plusmn 099 595 plusmn 024 532 plusmn 390 2121 plusmn 310 14750 plusmn 481 003 plusmn 001 565 plusmn 074
QUE 079 plusmn 008 455 plusmn 069 539 plusmn 086 490 plusmn 059 2024 plusmn 166 13064 plusmn 339 002 plusmn 000 455 plusmn 090
AA2 088 plusmn 004 524 plusmn 012 603 plusmn 056 511 plusmn 004 2046 plusmn 077 14446 plusmn 383 003 plusmn 001 392 plusmn 091
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Reducing LDL-C and TC can prevent the risk of CVDa leading cause of mortality worldwide [35] Appropriatelifestyle changes and pharmacologic approaches have bothdemonstrated their effectiveness in lowering LDL-C and TC[36] but the negative side effects of the pharmacologicalintervention have been a major setback Lifestyle changesto include decreased saturated fats and increased solublefibre in the diet weight loss and regular physical activity areprimary strategy for preventing CVD Regular consumptionof dietary supplements or functional foods that have demon-strated positive effects on plasma lipid values in randomisedplacebo-controlled clinical studies can also be considered aspart of this CVD prevention strategy [37] On this basiswe investigated the effects of methanol extract of AA onatherogenic indices and redox status of cellular system inhypercholesterolemic (HC) rats
The present study clearly shows that feeding rats onhigh cholesterol diets for nine weeks caused significantincrease in relative weight of heart and liver of the rats Thisobservation is consistent with the findings of Adaramoyeet al [16] and Yuji et al [38] However treatment withAA (100 and 200mgkg) and Questran significantly reducedthe relative weight of liver and heart of the HC rats Inthis study HC rats had high serum hepatic and cardiac
TC TG and LDL-C when compared to controls Similarobservations on hypercholesterolemic rats were observed byYuji et al [38] Adaramoye et al [39] and Kamesh andSumathi [40] Furthermore a decrease in serum HDL-Clevels was also observed in HC rats which actually reflectsthe lower cholesterol transports by HDL-C in blood fromperipheral tissues to liver for its metabolism and excretion[41] The elevated serum and tissues levels of TC TG andLDL-C and lower levels of HDL-C provide a high risk forthe development of atherosclerosis and other CVD [42] Inthe study extract of AA significantly decreased the levels ofTC TG and LDL-C and increased HDL-C in the HC ratsas compared to controls The lipoproteins especially LDL-Care involved in depositing TC and TG on walls of coronaryarteries and initiate the process of atherosclerotic plaques[43] Reduced serum and tissues levels of TC TG and LDL-cfound in HC rats treated with doses (100 and 200mgkg) ofAA are among the beneficial aspects of this current researchand proved the antiatherosclerotic potential of this extractThe crucial risk factor for CVD includes a low level of HDL-C and high level of LDL-C The association between a lowlevel of HDL-C and an increased risk of CVD has beenwell established through epidemiological and clinical studies[44] Since low level of HDL-C plays a direct role in the
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
2 Advances in Pharmacological Sciences
are used for the treatment of liver disorders hypertensionand diabetes [11 12] In vitro studies by Nwokocha et al[13] supported the folkloric use of this plant However littleinformation is available with respect to in vivo studies on itsethnomedicinal usesThis study was designed to evaluate theeffects of Artocarpus altilis on atherogenic indices and redoxstatus of cellular system of hypercholesterolemic rats
2 Materials and Methods
21 Chemicals Questran (Bristol-Myers Squibb HounslowUK) was purchased from a local chemist in IbadanNigeria Dietary cholesterol and thiobarbituric acid (TBA)were procured from Aldrich Chemical Co (MilwaukeeWI USA) Glutathione hydrogen peroxide 551015840-dithio-bis-2-nitrobenzoic acid (DNTB) and epinephrine were pur-chased from Sigma Chemical Co Saint Louis MO USATrichloroacetic acid (TCA) and thiobarbituric acid (TBA)were purchased from British Drug House (BDH) ChemicalLtd Poole UK Other reagents were of analytical grade andthe purest quality available
22 Collection and Extraction of Artocarpus altilis The stembark of Artocarpus altilis was collected in Ibadan (Oyo State)and authenticated at the BotanicalGarden of theUniversity ofIbadan The stem bark of Artocarpus altilis was air-dried andcrushed into fine powder The powdered part was extractedwith n-hexane and methanol using soxhlet extractor andthe extract was concentrated in vacuum at 40∘C with rotaryevaporator and water bath to dryness The yield of theextraction was 57
23 Determination of Total Phenolic Contents The totalphenolic content of the extract was determined using themethod of Singleton et al [14] with slight modificationsFolin-C reagent (1mL) was added to 1mL of extract orstandard After 3 minutes 1mL of 15 Na
2CO3was added
and the solution was made up to 5mL with distilled waterThe reaction mixture was kept in the dark for 90 minuteswith intermittent shaking or placed in a water bath at 40∘Cfor 20 minutes The absorbance was measured by a BeckmanDU (70) Spectrophotometer at 760 nm All experimentswere done in triplicate A standard curve was plotted withcatechin and the phenolic content expressed as CE (catechinequivalent) per mg dry weight of the extract
231 DPPHmdashRadical Scavenging Activity The radical scav-enging activity of the extract was measured as described byMensor et al [15] The stable 22-diphenyl-1-picrylhydrazyl(DPPH) radical was used for the determination of freeradical scavenging activities of the extracts A portion (1mL)of each of the different concentrations (10ndash1000 120583gmL) ofthe extracts or standard (catechin) was added to 1mL of1mM DPPH in methanol The mixtures were vortexed andincubated in a dark chamber for 30 minutes after which theabsorbance was measured at 517 nm against a DPPH controlcontaining only 1mL of methanol in place of the extract All
calculations were carried out in triplicates The inhibition ofDPPH was calculated as a percentage using the expression
119868 =119860control minus 119860 sample
119860controltimes 100 (1)
where 119868 is the percentage inhibition of the DPPH radical119860control is the absorbance of the control and 119860 sample is theabsorbance of the test compound
24 Animals Inbred male Wistar rats weighing between150 and 180 g were purchased from the animal house ofthe Department of Veterinary Physiology Biochemistry andPharmacology University of Ibadan Nigeria Animals werekept in ventilated cages at room temperature (28ndash30∘C) andmaintained on normal laboratory chow (Ladokun FeedsIbadan Nigeria) and water ad libitum Rats handling andtreatments conform to guidelines of the National Institute ofHealth (NIH publication 85-23 1985) for laboratory animalcare and use The study was approved by the Faculty ofBasic Medical Sciences University of Ibadan Animal EthicsCommittee
25 Study Design Thirty-five male rats were randomlydivided into seven groups of five rats each The first group(control) received drug vehicle (corn oil) the second group(HC) received dietary cholesterol at 30mg03mL [16]the third group (HC + AA1) received dietary cholesteroland Artocarpus altilis (100mgkg) the fourth group (HC+ AA2) received dietary cholesterol and Artocarpus altilis(200mgkg) the fifth group (HC + QUE) received dietarycholesterol and Questran (026 gkg) [16] the sixth group(QUE) received questran alone and the seventh group (AA)receivedArtocarpus altilis at a dose of 200mgkg bodyweight
26 Preparation of Tissues Rats were fasted overnight andsacrificed 24 hours after the last dose of drugs Liver andheart were quickly removed and washed in ice-cold 115KCl solution dried and weighed A section of liver andaorta samples were fixed in 10 formalin for histologicalexamination The remaining parts of liver and heart werehomogenized in 4 volumes of 50mM phosphate buffer pH74 and centrifuged at 10000 g for 15 minutes to obtain post-mitochondrial supernatant fraction (PMF) All procedureswere carried out at temperature of 0ndash4∘C
261 Preparation of Serum Blood was collected from theheart of the animals into plain centrifuge tubes and wasallowed to stand for 1 hour Serum was prepared by cen-trifugation at 3000 g for 15 minutes in a Beckman benchcentrifugeThe clear supernatant was used for the estimationof serum lipid profile and enzymes
27 Biochemical Assays Protein contents of the samples wereassayed by the method of Lowry et al [17] using bovineserum albumin as standard The activities of alanine andaspartate aminotransferases (ALT and AST) were assayed bythe combinedmethods ofMohun andCook [18] andReitman
Advances in Pharmacological Sciences 3
Table 1 Changes in the body weight and relative weight of organs of hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
Treatment Body weight (g) Weight of organs (g) Relative weight of organsInitial Final Liver Kidney Heart Liver Kidney Heart
Control 15100 plusmn 494 19400 plusmn 2302 538 plusmn 049 103 plusmn 011 053 plusmn 009 277 plusmn 075 053 plusmn 005 027 plusmn 004
HC 16000 plusmn 310 19800 plusmn 2903 689 plusmn 089 109 plusmn 014 069 plusmn 006 348 plusmn 023lowast055 plusmn 003 039 plusmn 002
lowast
HC + AA1 15900 plusmn 548 19600 plusmn 3715 512 plusmn 122 109 plusmn 020 055 plusmn 008 261 plusmn 013lowastlowast 056 plusmn 004 028 plusmn 003lowastlowast
HC + AA2 16700 plusmn 604 18500 plusmn 4500 520 plusmn 124 109 plusmn 020 052 plusmn 009 281 plusmn 015lowastlowast 059 plusmn 006 028 plusmn 004lowastlowast
HC + QUE 17300 plusmn 474 20250 plusmn 500 539 plusmn 105 119 plusmn 007 055 plusmn 001 266 plusmn 043lowastlowast 059 plusmn 005 027 plusmn 001lowastlowast
QUE 18800 plusmn 395 22500 plusmn 2041 506 plusmn 121 117 plusmn 005 051 plusmn 049 224 plusmn 041 052 plusmn 007 023 plusmn 004
AA2 19200 plusmn 834 23500 plusmn 2236 519 plusmn 055 121 plusmn 011 062 plusmn 006 221 plusmn 017 052 plusmn 004 026 plusmn 003
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
and Frankel [19] Serum total cholesterol level was assayedby the method of Richmond [20] The method involvedenzymatic hydrolysis and oxidation of cholesterol with theformation of quinoneimine (an indicator) from hydrogenperoxide and 4-aminoantipyrine in the presence of phenoland peroxideThe serum level of triglyceride was determinedby Jacobs and van Demark [21] and Koditschek and Umbreit[22] this was based on the hydrolysis of triglycerides with theformation of glycerol which is substrate for other enzymeswith the subsequent formation of hydrogen peroxide Thisthen reacts with 4-aminophenazone and 4-chlorophenol inthe presence of peroxidase to give quinoneimine which ismeasured spectrophotometrically at 500 nm
The lipoproteins (measured using the enzymatic col-orimetric method) very low-density lipoprotein (VLDL)and low-density lipoprotein (LDL) were precipitated by theaddition of phosphotungstic acid and magnesium chlorideAfter centrifugation at 3000 g for 10 minutes at 25∘C theclear supernatant containedHDL fraction whichwas assayedfor cholesterol with the Randox diagnostic kit The low-density lipoprotein (LDL) was calculated using the formulaof Friedewald et al [23] Lipid peroxidation level was assayedby the reaction between 2-thiobarbituric acid (TBA) andmalondialdehyde (MDA) an end product of lipid peroxidesas described by Buege and Aust [24]
The activity of lactate dehydrogenase (LDH) was deter-mined by the method of Zimmerman and Weinstein [25]while tissue superoxide dismutase (SOD) activity was mea-sured by the nitro blue tetrazolium (NBT) reduction methodof McCord and Fridovich [26] Catalase (CAT) activity wasassayed spectrophotometrically by measuring the rate ofdecomposition of hydrogen peroxide at 240 nm as describedby Aebi [27] Reduced glutathione level was measured by themethod of Beutler et al [28] this method is based on thedevelopment of a relatively stable (yellow) colour when 5101584051015840-dithiobis-(2-nitrobenzoic acid) (Ellmanrsquos reagent) is added tosulfhydryl compounds The chromophoric product resultingfrom the reaction of Ellmanrsquos reagent with the reducedglutathione (2-nitro-5-thiobenzoic acid) possesses a molarabsorption at 412 nm which is proportion to the level ofreduced glutathione in the test sample The glutathione per-oxidase (GPx) activity was assessed by themethod of Rotruck
et al [29] while glutathione-S-transferase (GST) activity wasdetermined according to Habig et al [30] the principle isbased on the fact that all of known GST demonstrates arelatively high activity with 1-chloro-24-dinitrobenzene asthe second substrate When this substance is conjugated withreduced glutathione its absorption maximum shifts to alonger wavelength 340 nm and the absorption increase at thiswavelength provides a direct measurement of the enzymaticreaction
271 Determination of Antiatherogenic Cardioprotective andCoronary Risk Indices Cardioprotective index (CPI) wasestimated in terms of HDL-C to LDL-C ratio [31 32] whereasantiatherogenic (AAI) and coronary risk indices (CRI) werecalculated by the following formulae [33 34]
AAI = 100 times [HDL-C][Total cholesterol minusHDL-C ]
CRI = Total cholesterolHDL-cholesterol
(2)
28 Histopathology of Tissues Tissues fixed in 10 formalinwere dehydrated in 95 ethanol and then cleared in xylenebefore embedded in paraffin Microsections (about 4 120583m)were prepared and stained with haematoxylin and eosin(HampE) dye and were examined under a light microscope by ahistopathologist who was ignorant of the treatment groups
29 Statistical Analysis All values were expressed as themean plusmn SD of five animals per group Data were analyzedusing one-way ANOVA followed by the post hoc Duncanmultiple range test for analysis of biochemical data usingSPSS (100) Values were considered statistically significant at119875 lt 005
3 Results
31 Phenolic and Flavonoids Contents and Effects of Arto-carpus altilis on Body Weight and Relative Weight of Organsof Hypercholesterolemic (HC) Rats In Table 1 there weresignificant increases (119875 lt 005) in the relative weight of liver
4 Advances in Pharmacological Sciences
Table 2 The total phenolic contents and scavenging activity ofArtocarpus altilis on 22-diphenyl-1-picrylhydrazyl radical (DPPH)in vitro
Conc Scavenging activity Phenolic content(120583gmL) Catechin AA (120583gCEmg)100 422 plusmn 44 227 plusmn 59 018 plusmn 002
300 476 plusmn 16 504 plusmn 31 039 plusmn 003
500 631 plusmn 51 511 plusmn 23 052 plusmn 007
750 678 plusmn 39 625 plusmn 85 068 plusmn 005
Data are expressed as mean plusmn SD (119899 = 4)AA Artocarpus altilis
and heart of HC rats when compared with the control whiletreatment with AA (100 and 200mgkg) significantly reducedthe relative weight of heart and liver of HC rats to values thatwere statistically similar (119875 gt 005) to the control Similarreductionwas obtained in questran-treatedHC ratsThe totalphenolic contents (TPC) of AA expressed in 120583g catechinequivalent per mg dry weight of the extract increased withincrease in concentration (Table 2) At 750 120583gmL the TPCof AA was 068 plusmn 005 120583gCEmg There were significant(119875 lt 005) and dose-dependent increases in scavengingactivity of AA on DPPH radicals (Table 2) At 100 120583gmLand 750 120583gmL the percentage DPPH radical scavengingactivities of AA were 422 and 678 respectively
32 Effects of Artocarpus altilis on Antioxidant Parametersand Marker Enzymes in Hypercholesterolemic (HC) RatsAdministration of dietary cholesterol significantly increased(119875 lt 005) serum hepatic and cardiac lipid peroxidation(LPO) products measured as thiobarbituric acid reactivesubstances (TBARS) by 265 83 and 80 respectively(Table 3) However treatment with AA completely amelio-rated dietary cholesterol-induced increase in LPO In HCrats the activities of hepatic and cardiac SOD and CATas well as cardiac GPx decreased significantly relative tothe control (Table 4) Specifically hepatic SOD and CATdecreased by 54and 45while cardiac SODCAT andGPxdecreased by 67 59 and 36 respectively Also activitiesof phase II and antioxidant enzyme (GST) in the liver of HCrats were significantly reduced when compared to controls(Figure 5) Administration of AA (200mgkg) reversed theadverse effect of high dietary cholesterol by normalizing theseenzymic antioxidant indices In HC rats serum ALT ASTand LDH were significantly increased by 23- 17- and 24-fold respectively while cardiac LDH activity was decreasedby 30-fold relative to controls (Table 5 and Figures 3 and4) However the observed elevations in the activities of theseserum enzymes in HC rats were reversed following treatmentwith AA and quetsran
33 Effects of Artocarpus altilis on the Lipid Profile of Hyperc-holesterolemic Rats Feeding rats on high dietary cholesterolfor nine consecutive weeks significantly (119875 lt 005) increasedthe serum hepatic and cardiac total cholesterol levels by110 70 and 85 respectively (Table 6 and Figures 1 and2) Furthermore serum hepatic and cardiac triglyceridesincreased by 68 96 and 62 while serum LDL-C
0
200
400
600
800
1000
1200
1400
1600
Con
c (m
gdL
)Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Heart total cholHeart trig
lowast
lowast
lowastlowastlowastlowast
lowastlowast
lowastlowastlowastlowast
Figure 1 Effects of methanol extract of Artocarpus altilis andQuestran on cardiac total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC Hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
increased by 79 respectively in HC rats relative to controlsIn addition HC rats had significantly lower HDL-C valueswhen compared to the control (Table 6) Administration ofAA at 200mgkg attenuated the elevated levels of these lipidindices to near normal in the tissues ofHC ratsTheprotectiveeffect of AA at 200mgkg seems better than the standardhypolipidemic drug (Questran) Furthermore AA increasedserum antiatherogenic index in HC rats while coronary riskindex was decreased (Table 7)
34 Effects of Artocarpus altilis on the Histology of Aorta andLiver Thehistology of liver slide showedmarked portal con-gestion severe periportal cellular infiltration bymononuclearcells and mild diffuse vacuolar degeneration of hepatocytes(Figure 6) while aorta from HC rats revealed large focal areaof myofibril necrosis with severe hemorrhages and fibrousconnective tissue laid down (Figure 7) Treatment withAA (200mgkg) reversed the adverse effect of high dietarycholesterol on the histological architecture of the aorta andliver of the rats The histological results further corroboratedthe biochemical findings indicating the beneficial effects ofAA in hypercholesterolemic rats
4 Discussion
It is generally known that elevation of serum LDL-C and totalcholesterol (TC) can lead to CVD especially atherosclerosis
Advances in Pharmacological Sciences 5
Table 3 Changes in the levels of lipid peroxidation in hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nineweeks
Treatments Liver(120583molMDAmg protein)
Heart(120583molMDAmg protein)
Serum(120583molMDAmg protein)
Control 006 plusmn 001 015 plusmn 002 132 plusmn 024
HC 011 plusmn 002lowast
027 plusmn 003lowast
482 plusmn 068lowast
HC + AA1 008 plusmn 003lowastlowast
013 plusmn 003lowastlowast
263 plusmn 056lowastlowast
HC + AA2 007 plusmn 002lowastlowast
014 plusmn 003lowastlowast
204 plusmn 035lowastlowast
HC + QUE 008 plusmn 002 015 plusmn 002 285 plusmn 077
QUE 005 plusmn 001 016 plusmn 007 182 plusmn 060
AA2 007 plusmn 001 015 plusmn 002 116 plusmn 033
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 4 Changes in the levels of hepatic and cardiac antioxidant parameters in hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
TreatmentLiver Heart
GSH GPx SOD CAT GSH GPx SOD CAT(mgg tissue) (Umg protein) (mgg tissue) (Umg protein)
Control 085 plusmn 015 555 plusmn 083 736 plusmn 101 557 plusmn 108 2106 plusmn 111 14574 plusmn 574 003 plusmn 001 537 plusmn 081
HC 073 plusmn 001 492 plusmn 008 340 plusmn 069lowast
304 plusmn 005lowast1925 plusmn 099 9282 plusmn 287
lowast
001 plusmn 001lowast
219 plusmn 065lowast
HC + AA1 092 plusmn 026 498 plusmn 006 726 plusmn 100lowastlowast 543 plusmn 171lowastlowast 1935 plusmn 085 10843 plusmn 287 001 plusmn 001 299 plusmn 074
HC + AA2 096 plusmn 025 573 plusmn 150 743 plusmn 071lowastlowast 555 plusmn 159lowastlowast 2038 plusmn 111 13747 plusmn 301lowastlowast 003 plusmn 000lowastlowast 403 plusmn 080lowastlowast
HC + QUE 085 plusmn 012 537 plusmn 099 595 plusmn 024 532 plusmn 390 2121 plusmn 310 14750 plusmn 481 003 plusmn 001 565 plusmn 074
QUE 079 plusmn 008 455 plusmn 069 539 plusmn 086 490 plusmn 059 2024 plusmn 166 13064 plusmn 339 002 plusmn 000 455 plusmn 090
AA2 088 plusmn 004 524 plusmn 012 603 plusmn 056 511 plusmn 004 2046 plusmn 077 14446 plusmn 383 003 plusmn 001 392 plusmn 091
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Reducing LDL-C and TC can prevent the risk of CVDa leading cause of mortality worldwide [35] Appropriatelifestyle changes and pharmacologic approaches have bothdemonstrated their effectiveness in lowering LDL-C and TC[36] but the negative side effects of the pharmacologicalintervention have been a major setback Lifestyle changesto include decreased saturated fats and increased solublefibre in the diet weight loss and regular physical activity areprimary strategy for preventing CVD Regular consumptionof dietary supplements or functional foods that have demon-strated positive effects on plasma lipid values in randomisedplacebo-controlled clinical studies can also be considered aspart of this CVD prevention strategy [37] On this basiswe investigated the effects of methanol extract of AA onatherogenic indices and redox status of cellular system inhypercholesterolemic (HC) rats
The present study clearly shows that feeding rats onhigh cholesterol diets for nine weeks caused significantincrease in relative weight of heart and liver of the rats Thisobservation is consistent with the findings of Adaramoyeet al [16] and Yuji et al [38] However treatment withAA (100 and 200mgkg) and Questran significantly reducedthe relative weight of liver and heart of the HC rats Inthis study HC rats had high serum hepatic and cardiac
TC TG and LDL-C when compared to controls Similarobservations on hypercholesterolemic rats were observed byYuji et al [38] Adaramoye et al [39] and Kamesh andSumathi [40] Furthermore a decrease in serum HDL-Clevels was also observed in HC rats which actually reflectsthe lower cholesterol transports by HDL-C in blood fromperipheral tissues to liver for its metabolism and excretion[41] The elevated serum and tissues levels of TC TG andLDL-C and lower levels of HDL-C provide a high risk forthe development of atherosclerosis and other CVD [42] Inthe study extract of AA significantly decreased the levels ofTC TG and LDL-C and increased HDL-C in the HC ratsas compared to controls The lipoproteins especially LDL-Care involved in depositing TC and TG on walls of coronaryarteries and initiate the process of atherosclerotic plaques[43] Reduced serum and tissues levels of TC TG and LDL-cfound in HC rats treated with doses (100 and 200mgkg) ofAA are among the beneficial aspects of this current researchand proved the antiatherosclerotic potential of this extractThe crucial risk factor for CVD includes a low level of HDL-C and high level of LDL-C The association between a lowlevel of HDL-C and an increased risk of CVD has beenwell established through epidemiological and clinical studies[44] Since low level of HDL-C plays a direct role in the
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
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StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
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MEDIATORSINFLAMMATION
of
Advances in Pharmacological Sciences 3
Table 1 Changes in the body weight and relative weight of organs of hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
Treatment Body weight (g) Weight of organs (g) Relative weight of organsInitial Final Liver Kidney Heart Liver Kidney Heart
Control 15100 plusmn 494 19400 plusmn 2302 538 plusmn 049 103 plusmn 011 053 plusmn 009 277 plusmn 075 053 plusmn 005 027 plusmn 004
HC 16000 plusmn 310 19800 plusmn 2903 689 plusmn 089 109 plusmn 014 069 plusmn 006 348 plusmn 023lowast055 plusmn 003 039 plusmn 002
lowast
HC + AA1 15900 plusmn 548 19600 plusmn 3715 512 plusmn 122 109 plusmn 020 055 plusmn 008 261 plusmn 013lowastlowast 056 plusmn 004 028 plusmn 003lowastlowast
HC + AA2 16700 plusmn 604 18500 plusmn 4500 520 plusmn 124 109 plusmn 020 052 plusmn 009 281 plusmn 015lowastlowast 059 plusmn 006 028 plusmn 004lowastlowast
HC + QUE 17300 plusmn 474 20250 plusmn 500 539 plusmn 105 119 plusmn 007 055 plusmn 001 266 plusmn 043lowastlowast 059 plusmn 005 027 plusmn 001lowastlowast
QUE 18800 plusmn 395 22500 plusmn 2041 506 plusmn 121 117 plusmn 005 051 plusmn 049 224 plusmn 041 052 plusmn 007 023 plusmn 004
AA2 19200 plusmn 834 23500 plusmn 2236 519 plusmn 055 121 plusmn 011 062 plusmn 006 221 plusmn 017 052 plusmn 004 026 plusmn 003
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
and Frankel [19] Serum total cholesterol level was assayedby the method of Richmond [20] The method involvedenzymatic hydrolysis and oxidation of cholesterol with theformation of quinoneimine (an indicator) from hydrogenperoxide and 4-aminoantipyrine in the presence of phenoland peroxideThe serum level of triglyceride was determinedby Jacobs and van Demark [21] and Koditschek and Umbreit[22] this was based on the hydrolysis of triglycerides with theformation of glycerol which is substrate for other enzymeswith the subsequent formation of hydrogen peroxide Thisthen reacts with 4-aminophenazone and 4-chlorophenol inthe presence of peroxidase to give quinoneimine which ismeasured spectrophotometrically at 500 nm
The lipoproteins (measured using the enzymatic col-orimetric method) very low-density lipoprotein (VLDL)and low-density lipoprotein (LDL) were precipitated by theaddition of phosphotungstic acid and magnesium chlorideAfter centrifugation at 3000 g for 10 minutes at 25∘C theclear supernatant containedHDL fraction whichwas assayedfor cholesterol with the Randox diagnostic kit The low-density lipoprotein (LDL) was calculated using the formulaof Friedewald et al [23] Lipid peroxidation level was assayedby the reaction between 2-thiobarbituric acid (TBA) andmalondialdehyde (MDA) an end product of lipid peroxidesas described by Buege and Aust [24]
The activity of lactate dehydrogenase (LDH) was deter-mined by the method of Zimmerman and Weinstein [25]while tissue superoxide dismutase (SOD) activity was mea-sured by the nitro blue tetrazolium (NBT) reduction methodof McCord and Fridovich [26] Catalase (CAT) activity wasassayed spectrophotometrically by measuring the rate ofdecomposition of hydrogen peroxide at 240 nm as describedby Aebi [27] Reduced glutathione level was measured by themethod of Beutler et al [28] this method is based on thedevelopment of a relatively stable (yellow) colour when 5101584051015840-dithiobis-(2-nitrobenzoic acid) (Ellmanrsquos reagent) is added tosulfhydryl compounds The chromophoric product resultingfrom the reaction of Ellmanrsquos reagent with the reducedglutathione (2-nitro-5-thiobenzoic acid) possesses a molarabsorption at 412 nm which is proportion to the level ofreduced glutathione in the test sample The glutathione per-oxidase (GPx) activity was assessed by themethod of Rotruck
et al [29] while glutathione-S-transferase (GST) activity wasdetermined according to Habig et al [30] the principle isbased on the fact that all of known GST demonstrates arelatively high activity with 1-chloro-24-dinitrobenzene asthe second substrate When this substance is conjugated withreduced glutathione its absorption maximum shifts to alonger wavelength 340 nm and the absorption increase at thiswavelength provides a direct measurement of the enzymaticreaction
271 Determination of Antiatherogenic Cardioprotective andCoronary Risk Indices Cardioprotective index (CPI) wasestimated in terms of HDL-C to LDL-C ratio [31 32] whereasantiatherogenic (AAI) and coronary risk indices (CRI) werecalculated by the following formulae [33 34]
AAI = 100 times [HDL-C][Total cholesterol minusHDL-C ]
CRI = Total cholesterolHDL-cholesterol
(2)
28 Histopathology of Tissues Tissues fixed in 10 formalinwere dehydrated in 95 ethanol and then cleared in xylenebefore embedded in paraffin Microsections (about 4 120583m)were prepared and stained with haematoxylin and eosin(HampE) dye and were examined under a light microscope by ahistopathologist who was ignorant of the treatment groups
29 Statistical Analysis All values were expressed as themean plusmn SD of five animals per group Data were analyzedusing one-way ANOVA followed by the post hoc Duncanmultiple range test for analysis of biochemical data usingSPSS (100) Values were considered statistically significant at119875 lt 005
3 Results
31 Phenolic and Flavonoids Contents and Effects of Arto-carpus altilis on Body Weight and Relative Weight of Organsof Hypercholesterolemic (HC) Rats In Table 1 there weresignificant increases (119875 lt 005) in the relative weight of liver
4 Advances in Pharmacological Sciences
Table 2 The total phenolic contents and scavenging activity ofArtocarpus altilis on 22-diphenyl-1-picrylhydrazyl radical (DPPH)in vitro
Conc Scavenging activity Phenolic content(120583gmL) Catechin AA (120583gCEmg)100 422 plusmn 44 227 plusmn 59 018 plusmn 002
300 476 plusmn 16 504 plusmn 31 039 plusmn 003
500 631 plusmn 51 511 plusmn 23 052 plusmn 007
750 678 plusmn 39 625 plusmn 85 068 plusmn 005
Data are expressed as mean plusmn SD (119899 = 4)AA Artocarpus altilis
and heart of HC rats when compared with the control whiletreatment with AA (100 and 200mgkg) significantly reducedthe relative weight of heart and liver of HC rats to values thatwere statistically similar (119875 gt 005) to the control Similarreductionwas obtained in questran-treatedHC ratsThe totalphenolic contents (TPC) of AA expressed in 120583g catechinequivalent per mg dry weight of the extract increased withincrease in concentration (Table 2) At 750 120583gmL the TPCof AA was 068 plusmn 005 120583gCEmg There were significant(119875 lt 005) and dose-dependent increases in scavengingactivity of AA on DPPH radicals (Table 2) At 100 120583gmLand 750 120583gmL the percentage DPPH radical scavengingactivities of AA were 422 and 678 respectively
32 Effects of Artocarpus altilis on Antioxidant Parametersand Marker Enzymes in Hypercholesterolemic (HC) RatsAdministration of dietary cholesterol significantly increased(119875 lt 005) serum hepatic and cardiac lipid peroxidation(LPO) products measured as thiobarbituric acid reactivesubstances (TBARS) by 265 83 and 80 respectively(Table 3) However treatment with AA completely amelio-rated dietary cholesterol-induced increase in LPO In HCrats the activities of hepatic and cardiac SOD and CATas well as cardiac GPx decreased significantly relative tothe control (Table 4) Specifically hepatic SOD and CATdecreased by 54and 45while cardiac SODCAT andGPxdecreased by 67 59 and 36 respectively Also activitiesof phase II and antioxidant enzyme (GST) in the liver of HCrats were significantly reduced when compared to controls(Figure 5) Administration of AA (200mgkg) reversed theadverse effect of high dietary cholesterol by normalizing theseenzymic antioxidant indices In HC rats serum ALT ASTand LDH were significantly increased by 23- 17- and 24-fold respectively while cardiac LDH activity was decreasedby 30-fold relative to controls (Table 5 and Figures 3 and4) However the observed elevations in the activities of theseserum enzymes in HC rats were reversed following treatmentwith AA and quetsran
33 Effects of Artocarpus altilis on the Lipid Profile of Hyperc-holesterolemic Rats Feeding rats on high dietary cholesterolfor nine consecutive weeks significantly (119875 lt 005) increasedthe serum hepatic and cardiac total cholesterol levels by110 70 and 85 respectively (Table 6 and Figures 1 and2) Furthermore serum hepatic and cardiac triglyceridesincreased by 68 96 and 62 while serum LDL-C
0
200
400
600
800
1000
1200
1400
1600
Con
c (m
gdL
)Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Heart total cholHeart trig
lowast
lowast
lowastlowastlowastlowast
lowastlowast
lowastlowastlowastlowast
Figure 1 Effects of methanol extract of Artocarpus altilis andQuestran on cardiac total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC Hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
increased by 79 respectively in HC rats relative to controlsIn addition HC rats had significantly lower HDL-C valueswhen compared to the control (Table 6) Administration ofAA at 200mgkg attenuated the elevated levels of these lipidindices to near normal in the tissues ofHC ratsTheprotectiveeffect of AA at 200mgkg seems better than the standardhypolipidemic drug (Questran) Furthermore AA increasedserum antiatherogenic index in HC rats while coronary riskindex was decreased (Table 7)
34 Effects of Artocarpus altilis on the Histology of Aorta andLiver Thehistology of liver slide showedmarked portal con-gestion severe periportal cellular infiltration bymononuclearcells and mild diffuse vacuolar degeneration of hepatocytes(Figure 6) while aorta from HC rats revealed large focal areaof myofibril necrosis with severe hemorrhages and fibrousconnective tissue laid down (Figure 7) Treatment withAA (200mgkg) reversed the adverse effect of high dietarycholesterol on the histological architecture of the aorta andliver of the rats The histological results further corroboratedthe biochemical findings indicating the beneficial effects ofAA in hypercholesterolemic rats
4 Discussion
It is generally known that elevation of serum LDL-C and totalcholesterol (TC) can lead to CVD especially atherosclerosis
Advances in Pharmacological Sciences 5
Table 3 Changes in the levels of lipid peroxidation in hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nineweeks
Treatments Liver(120583molMDAmg protein)
Heart(120583molMDAmg protein)
Serum(120583molMDAmg protein)
Control 006 plusmn 001 015 plusmn 002 132 plusmn 024
HC 011 plusmn 002lowast
027 plusmn 003lowast
482 plusmn 068lowast
HC + AA1 008 plusmn 003lowastlowast
013 plusmn 003lowastlowast
263 plusmn 056lowastlowast
HC + AA2 007 plusmn 002lowastlowast
014 plusmn 003lowastlowast
204 plusmn 035lowastlowast
HC + QUE 008 plusmn 002 015 plusmn 002 285 plusmn 077
QUE 005 plusmn 001 016 plusmn 007 182 plusmn 060
AA2 007 plusmn 001 015 plusmn 002 116 plusmn 033
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 4 Changes in the levels of hepatic and cardiac antioxidant parameters in hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
TreatmentLiver Heart
GSH GPx SOD CAT GSH GPx SOD CAT(mgg tissue) (Umg protein) (mgg tissue) (Umg protein)
Control 085 plusmn 015 555 plusmn 083 736 plusmn 101 557 plusmn 108 2106 plusmn 111 14574 plusmn 574 003 plusmn 001 537 plusmn 081
HC 073 plusmn 001 492 plusmn 008 340 plusmn 069lowast
304 plusmn 005lowast1925 plusmn 099 9282 plusmn 287
lowast
001 plusmn 001lowast
219 plusmn 065lowast
HC + AA1 092 plusmn 026 498 plusmn 006 726 plusmn 100lowastlowast 543 plusmn 171lowastlowast 1935 plusmn 085 10843 plusmn 287 001 plusmn 001 299 plusmn 074
HC + AA2 096 plusmn 025 573 plusmn 150 743 plusmn 071lowastlowast 555 plusmn 159lowastlowast 2038 plusmn 111 13747 plusmn 301lowastlowast 003 plusmn 000lowastlowast 403 plusmn 080lowastlowast
HC + QUE 085 plusmn 012 537 plusmn 099 595 plusmn 024 532 plusmn 390 2121 plusmn 310 14750 plusmn 481 003 plusmn 001 565 plusmn 074
QUE 079 plusmn 008 455 plusmn 069 539 plusmn 086 490 plusmn 059 2024 plusmn 166 13064 plusmn 339 002 plusmn 000 455 plusmn 090
AA2 088 plusmn 004 524 plusmn 012 603 plusmn 056 511 plusmn 004 2046 plusmn 077 14446 plusmn 383 003 plusmn 001 392 plusmn 091
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Reducing LDL-C and TC can prevent the risk of CVDa leading cause of mortality worldwide [35] Appropriatelifestyle changes and pharmacologic approaches have bothdemonstrated their effectiveness in lowering LDL-C and TC[36] but the negative side effects of the pharmacologicalintervention have been a major setback Lifestyle changesto include decreased saturated fats and increased solublefibre in the diet weight loss and regular physical activity areprimary strategy for preventing CVD Regular consumptionof dietary supplements or functional foods that have demon-strated positive effects on plasma lipid values in randomisedplacebo-controlled clinical studies can also be considered aspart of this CVD prevention strategy [37] On this basiswe investigated the effects of methanol extract of AA onatherogenic indices and redox status of cellular system inhypercholesterolemic (HC) rats
The present study clearly shows that feeding rats onhigh cholesterol diets for nine weeks caused significantincrease in relative weight of heart and liver of the rats Thisobservation is consistent with the findings of Adaramoyeet al [16] and Yuji et al [38] However treatment withAA (100 and 200mgkg) and Questran significantly reducedthe relative weight of liver and heart of the HC rats Inthis study HC rats had high serum hepatic and cardiac
TC TG and LDL-C when compared to controls Similarobservations on hypercholesterolemic rats were observed byYuji et al [38] Adaramoye et al [39] and Kamesh andSumathi [40] Furthermore a decrease in serum HDL-Clevels was also observed in HC rats which actually reflectsthe lower cholesterol transports by HDL-C in blood fromperipheral tissues to liver for its metabolism and excretion[41] The elevated serum and tissues levels of TC TG andLDL-C and lower levels of HDL-C provide a high risk forthe development of atherosclerosis and other CVD [42] Inthe study extract of AA significantly decreased the levels ofTC TG and LDL-C and increased HDL-C in the HC ratsas compared to controls The lipoproteins especially LDL-Care involved in depositing TC and TG on walls of coronaryarteries and initiate the process of atherosclerotic plaques[43] Reduced serum and tissues levels of TC TG and LDL-cfound in HC rats treated with doses (100 and 200mgkg) ofAA are among the beneficial aspects of this current researchand proved the antiatherosclerotic potential of this extractThe crucial risk factor for CVD includes a low level of HDL-C and high level of LDL-C The association between a lowlevel of HDL-C and an increased risk of CVD has beenwell established through epidemiological and clinical studies[44] Since low level of HDL-C plays a direct role in the
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
4 Advances in Pharmacological Sciences
Table 2 The total phenolic contents and scavenging activity ofArtocarpus altilis on 22-diphenyl-1-picrylhydrazyl radical (DPPH)in vitro
Conc Scavenging activity Phenolic content(120583gmL) Catechin AA (120583gCEmg)100 422 plusmn 44 227 plusmn 59 018 plusmn 002
300 476 plusmn 16 504 plusmn 31 039 plusmn 003
500 631 plusmn 51 511 plusmn 23 052 plusmn 007
750 678 plusmn 39 625 plusmn 85 068 plusmn 005
Data are expressed as mean plusmn SD (119899 = 4)AA Artocarpus altilis
and heart of HC rats when compared with the control whiletreatment with AA (100 and 200mgkg) significantly reducedthe relative weight of heart and liver of HC rats to values thatwere statistically similar (119875 gt 005) to the control Similarreductionwas obtained in questran-treatedHC ratsThe totalphenolic contents (TPC) of AA expressed in 120583g catechinequivalent per mg dry weight of the extract increased withincrease in concentration (Table 2) At 750 120583gmL the TPCof AA was 068 plusmn 005 120583gCEmg There were significant(119875 lt 005) and dose-dependent increases in scavengingactivity of AA on DPPH radicals (Table 2) At 100 120583gmLand 750 120583gmL the percentage DPPH radical scavengingactivities of AA were 422 and 678 respectively
32 Effects of Artocarpus altilis on Antioxidant Parametersand Marker Enzymes in Hypercholesterolemic (HC) RatsAdministration of dietary cholesterol significantly increased(119875 lt 005) serum hepatic and cardiac lipid peroxidation(LPO) products measured as thiobarbituric acid reactivesubstances (TBARS) by 265 83 and 80 respectively(Table 3) However treatment with AA completely amelio-rated dietary cholesterol-induced increase in LPO In HCrats the activities of hepatic and cardiac SOD and CATas well as cardiac GPx decreased significantly relative tothe control (Table 4) Specifically hepatic SOD and CATdecreased by 54and 45while cardiac SODCAT andGPxdecreased by 67 59 and 36 respectively Also activitiesof phase II and antioxidant enzyme (GST) in the liver of HCrats were significantly reduced when compared to controls(Figure 5) Administration of AA (200mgkg) reversed theadverse effect of high dietary cholesterol by normalizing theseenzymic antioxidant indices In HC rats serum ALT ASTand LDH were significantly increased by 23- 17- and 24-fold respectively while cardiac LDH activity was decreasedby 30-fold relative to controls (Table 5 and Figures 3 and4) However the observed elevations in the activities of theseserum enzymes in HC rats were reversed following treatmentwith AA and quetsran
33 Effects of Artocarpus altilis on the Lipid Profile of Hyperc-holesterolemic Rats Feeding rats on high dietary cholesterolfor nine consecutive weeks significantly (119875 lt 005) increasedthe serum hepatic and cardiac total cholesterol levels by110 70 and 85 respectively (Table 6 and Figures 1 and2) Furthermore serum hepatic and cardiac triglyceridesincreased by 68 96 and 62 while serum LDL-C
0
200
400
600
800
1000
1200
1400
1600
Con
c (m
gdL
)Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Heart total cholHeart trig
lowast
lowast
lowastlowastlowastlowast
lowastlowast
lowastlowastlowastlowast
Figure 1 Effects of methanol extract of Artocarpus altilis andQuestran on cardiac total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC Hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
increased by 79 respectively in HC rats relative to controlsIn addition HC rats had significantly lower HDL-C valueswhen compared to the control (Table 6) Administration ofAA at 200mgkg attenuated the elevated levels of these lipidindices to near normal in the tissues ofHC ratsTheprotectiveeffect of AA at 200mgkg seems better than the standardhypolipidemic drug (Questran) Furthermore AA increasedserum antiatherogenic index in HC rats while coronary riskindex was decreased (Table 7)
34 Effects of Artocarpus altilis on the Histology of Aorta andLiver Thehistology of liver slide showedmarked portal con-gestion severe periportal cellular infiltration bymononuclearcells and mild diffuse vacuolar degeneration of hepatocytes(Figure 6) while aorta from HC rats revealed large focal areaof myofibril necrosis with severe hemorrhages and fibrousconnective tissue laid down (Figure 7) Treatment withAA (200mgkg) reversed the adverse effect of high dietarycholesterol on the histological architecture of the aorta andliver of the rats The histological results further corroboratedthe biochemical findings indicating the beneficial effects ofAA in hypercholesterolemic rats
4 Discussion
It is generally known that elevation of serum LDL-C and totalcholesterol (TC) can lead to CVD especially atherosclerosis
Advances in Pharmacological Sciences 5
Table 3 Changes in the levels of lipid peroxidation in hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nineweeks
Treatments Liver(120583molMDAmg protein)
Heart(120583molMDAmg protein)
Serum(120583molMDAmg protein)
Control 006 plusmn 001 015 plusmn 002 132 plusmn 024
HC 011 plusmn 002lowast
027 plusmn 003lowast
482 plusmn 068lowast
HC + AA1 008 plusmn 003lowastlowast
013 plusmn 003lowastlowast
263 plusmn 056lowastlowast
HC + AA2 007 plusmn 002lowastlowast
014 plusmn 003lowastlowast
204 plusmn 035lowastlowast
HC + QUE 008 plusmn 002 015 plusmn 002 285 plusmn 077
QUE 005 plusmn 001 016 plusmn 007 182 plusmn 060
AA2 007 plusmn 001 015 plusmn 002 116 plusmn 033
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 4 Changes in the levels of hepatic and cardiac antioxidant parameters in hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
TreatmentLiver Heart
GSH GPx SOD CAT GSH GPx SOD CAT(mgg tissue) (Umg protein) (mgg tissue) (Umg protein)
Control 085 plusmn 015 555 plusmn 083 736 plusmn 101 557 plusmn 108 2106 plusmn 111 14574 plusmn 574 003 plusmn 001 537 plusmn 081
HC 073 plusmn 001 492 plusmn 008 340 plusmn 069lowast
304 plusmn 005lowast1925 plusmn 099 9282 plusmn 287
lowast
001 plusmn 001lowast
219 plusmn 065lowast
HC + AA1 092 plusmn 026 498 plusmn 006 726 plusmn 100lowastlowast 543 plusmn 171lowastlowast 1935 plusmn 085 10843 plusmn 287 001 plusmn 001 299 plusmn 074
HC + AA2 096 plusmn 025 573 plusmn 150 743 plusmn 071lowastlowast 555 plusmn 159lowastlowast 2038 plusmn 111 13747 plusmn 301lowastlowast 003 plusmn 000lowastlowast 403 plusmn 080lowastlowast
HC + QUE 085 plusmn 012 537 plusmn 099 595 plusmn 024 532 plusmn 390 2121 plusmn 310 14750 plusmn 481 003 plusmn 001 565 plusmn 074
QUE 079 plusmn 008 455 plusmn 069 539 plusmn 086 490 plusmn 059 2024 plusmn 166 13064 plusmn 339 002 plusmn 000 455 plusmn 090
AA2 088 plusmn 004 524 plusmn 012 603 plusmn 056 511 plusmn 004 2046 plusmn 077 14446 plusmn 383 003 plusmn 001 392 plusmn 091
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Reducing LDL-C and TC can prevent the risk of CVDa leading cause of mortality worldwide [35] Appropriatelifestyle changes and pharmacologic approaches have bothdemonstrated their effectiveness in lowering LDL-C and TC[36] but the negative side effects of the pharmacologicalintervention have been a major setback Lifestyle changesto include decreased saturated fats and increased solublefibre in the diet weight loss and regular physical activity areprimary strategy for preventing CVD Regular consumptionof dietary supplements or functional foods that have demon-strated positive effects on plasma lipid values in randomisedplacebo-controlled clinical studies can also be considered aspart of this CVD prevention strategy [37] On this basiswe investigated the effects of methanol extract of AA onatherogenic indices and redox status of cellular system inhypercholesterolemic (HC) rats
The present study clearly shows that feeding rats onhigh cholesterol diets for nine weeks caused significantincrease in relative weight of heart and liver of the rats Thisobservation is consistent with the findings of Adaramoyeet al [16] and Yuji et al [38] However treatment withAA (100 and 200mgkg) and Questran significantly reducedthe relative weight of liver and heart of the HC rats Inthis study HC rats had high serum hepatic and cardiac
TC TG and LDL-C when compared to controls Similarobservations on hypercholesterolemic rats were observed byYuji et al [38] Adaramoye et al [39] and Kamesh andSumathi [40] Furthermore a decrease in serum HDL-Clevels was also observed in HC rats which actually reflectsthe lower cholesterol transports by HDL-C in blood fromperipheral tissues to liver for its metabolism and excretion[41] The elevated serum and tissues levels of TC TG andLDL-C and lower levels of HDL-C provide a high risk forthe development of atherosclerosis and other CVD [42] Inthe study extract of AA significantly decreased the levels ofTC TG and LDL-C and increased HDL-C in the HC ratsas compared to controls The lipoproteins especially LDL-Care involved in depositing TC and TG on walls of coronaryarteries and initiate the process of atherosclerotic plaques[43] Reduced serum and tissues levels of TC TG and LDL-cfound in HC rats treated with doses (100 and 200mgkg) ofAA are among the beneficial aspects of this current researchand proved the antiatherosclerotic potential of this extractThe crucial risk factor for CVD includes a low level of HDL-C and high level of LDL-C The association between a lowlevel of HDL-C and an increased risk of CVD has beenwell established through epidemiological and clinical studies[44] Since low level of HDL-C plays a direct role in the
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
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StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
Advances in Pharmacological Sciences 5
Table 3 Changes in the levels of lipid peroxidation in hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nineweeks
Treatments Liver(120583molMDAmg protein)
Heart(120583molMDAmg protein)
Serum(120583molMDAmg protein)
Control 006 plusmn 001 015 plusmn 002 132 plusmn 024
HC 011 plusmn 002lowast
027 plusmn 003lowast
482 plusmn 068lowast
HC + AA1 008 plusmn 003lowastlowast
013 plusmn 003lowastlowast
263 plusmn 056lowastlowast
HC + AA2 007 plusmn 002lowastlowast
014 plusmn 003lowastlowast
204 plusmn 035lowastlowast
HC + QUE 008 plusmn 002 015 plusmn 002 285 plusmn 077
QUE 005 plusmn 001 016 plusmn 007 182 plusmn 060
AA2 007 plusmn 001 015 plusmn 002 116 plusmn 033
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 4 Changes in the levels of hepatic and cardiac antioxidant parameters in hypercholesterolemic rats treated with methanol extract ofArtocarpus altilis for nine weeks
TreatmentLiver Heart
GSH GPx SOD CAT GSH GPx SOD CAT(mgg tissue) (Umg protein) (mgg tissue) (Umg protein)
Control 085 plusmn 015 555 plusmn 083 736 plusmn 101 557 plusmn 108 2106 plusmn 111 14574 plusmn 574 003 plusmn 001 537 plusmn 081
HC 073 plusmn 001 492 plusmn 008 340 plusmn 069lowast
304 plusmn 005lowast1925 plusmn 099 9282 plusmn 287
lowast
001 plusmn 001lowast
219 plusmn 065lowast
HC + AA1 092 plusmn 026 498 plusmn 006 726 plusmn 100lowastlowast 543 plusmn 171lowastlowast 1935 plusmn 085 10843 plusmn 287 001 plusmn 001 299 plusmn 074
HC + AA2 096 plusmn 025 573 plusmn 150 743 plusmn 071lowastlowast 555 plusmn 159lowastlowast 2038 plusmn 111 13747 plusmn 301lowastlowast 003 plusmn 000lowastlowast 403 plusmn 080lowastlowast
HC + QUE 085 plusmn 012 537 plusmn 099 595 plusmn 024 532 plusmn 390 2121 plusmn 310 14750 plusmn 481 003 plusmn 001 565 plusmn 074
QUE 079 plusmn 008 455 plusmn 069 539 plusmn 086 490 plusmn 059 2024 plusmn 166 13064 plusmn 339 002 plusmn 000 455 plusmn 090
AA2 088 plusmn 004 524 plusmn 012 603 plusmn 056 511 plusmn 004 2046 plusmn 077 14446 plusmn 383 003 plusmn 001 392 plusmn 091
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Reducing LDL-C and TC can prevent the risk of CVDa leading cause of mortality worldwide [35] Appropriatelifestyle changes and pharmacologic approaches have bothdemonstrated their effectiveness in lowering LDL-C and TC[36] but the negative side effects of the pharmacologicalintervention have been a major setback Lifestyle changesto include decreased saturated fats and increased solublefibre in the diet weight loss and regular physical activity areprimary strategy for preventing CVD Regular consumptionof dietary supplements or functional foods that have demon-strated positive effects on plasma lipid values in randomisedplacebo-controlled clinical studies can also be considered aspart of this CVD prevention strategy [37] On this basiswe investigated the effects of methanol extract of AA onatherogenic indices and redox status of cellular system inhypercholesterolemic (HC) rats
The present study clearly shows that feeding rats onhigh cholesterol diets for nine weeks caused significantincrease in relative weight of heart and liver of the rats Thisobservation is consistent with the findings of Adaramoyeet al [16] and Yuji et al [38] However treatment withAA (100 and 200mgkg) and Questran significantly reducedthe relative weight of liver and heart of the HC rats Inthis study HC rats had high serum hepatic and cardiac
TC TG and LDL-C when compared to controls Similarobservations on hypercholesterolemic rats were observed byYuji et al [38] Adaramoye et al [39] and Kamesh andSumathi [40] Furthermore a decrease in serum HDL-Clevels was also observed in HC rats which actually reflectsthe lower cholesterol transports by HDL-C in blood fromperipheral tissues to liver for its metabolism and excretion[41] The elevated serum and tissues levels of TC TG andLDL-C and lower levels of HDL-C provide a high risk forthe development of atherosclerosis and other CVD [42] Inthe study extract of AA significantly decreased the levels ofTC TG and LDL-C and increased HDL-C in the HC ratsas compared to controls The lipoproteins especially LDL-Care involved in depositing TC and TG on walls of coronaryarteries and initiate the process of atherosclerotic plaques[43] Reduced serum and tissues levels of TC TG and LDL-cfound in HC rats treated with doses (100 and 200mgkg) ofAA are among the beneficial aspects of this current researchand proved the antiatherosclerotic potential of this extractThe crucial risk factor for CVD includes a low level of HDL-C and high level of LDL-C The association between a lowlevel of HDL-C and an increased risk of CVD has beenwell established through epidemiological and clinical studies[44] Since low level of HDL-C plays a direct role in the
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
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ToxinsJournal of
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StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
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Medicinal ChemistryInternational Journal of
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AddictionJournal of
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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Pharmaceutics
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MEDIATORSINFLAMMATION
of
6 Advances in Pharmacological Sciences
Table 5 Changes in the activities of serum hepatic and cardiac alanine and aspartate aminotransferases in hypercholesterolemic rats treatedwith methanol extract of Artocarpus altilis for nine weeks
Treatments Liver (UL) Heart (UL) Serum (UL)AST ALT AST ALT AST ALT
Control 6106 plusmn 163 702 plusmn 54 5522 plusmn 242 3173 plusmn 123 2180 plusmn 180 520 plusmn 76
HC 6625 plusmn 193 727 plusmn 95 5868 plusmn 212 3302 plusmn 195 3628 plusmn 110lowast
1218 plusmn 105lowast
HC + AA1 6428 plusmn 137 694 plusmn 88 5494 plusmn 178 3410 plusmn 150 2438 plusmn 170lowastlowast
733 plusmn 43lowastlowast
HC + AA2 6413 plusmn 195 6793 plusmn 72 5712 plusmn 253 3094 plusmn 156 2587 plusmn 130lowastlowast
708 plusmn 48lowastlowast
HC + QUE 6422 plusmn 150 669 plusmn 018 5587 plusmn 182 282 plusmn 248 2635 plusmn 110lowastlowast
635 plusmn 85
QUE 6175 plusmn 153 693 plusmn 105 5691 plusmn 1354 2740 plusmn 113 2110 plusmn 96 604 plusmn 76
AA2 6170 plusmn 106 670 plusmn 79 5330 plusmn 225 3122 plusmn 174 2294 plusmn 165 640 plusmn 66
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
Table 6 Changes in serum lipid profile of hypercholesterolemic rats treated with methanol extract of Artocarpus altilis for nine weeks
Treatment Total chol Triglyceride(mgdL) LDL-C HDL-C
Control 32507 plusmn 771 41651 plusmn 2492 21140 plusmn 903 24213 plusmn 2955
HC 68081 plusmn 1642lowast
70177 plusmn 1863lowast
37655 plusmn 1371lowast
15039 plusmn 3249lowast
HC + AA1 51119 plusmn 1399lowastlowast
52144 plusmn 1513lowastlowast
36604 plusmn 1421 11015 plusmn 1400
HC + AA2 46407 plusmn 1665lowastlowast
48488 plusmn 1103lowastlowast
25739 plusmn 1650lowastlowast
22852 plusmn 1667lowastlowast
HC + QUE 37871 plusmn 1259 45246 plusmn 1317 30828 plusmn 1095 22301 plusmn 1467
QUE 41133 plusmn 1672 40951 plusmn 1408 29872 plusmn 1619 17619 plusmn 5172
AA2 34554 plusmn 1101 43869 plusmn 1863 22818 plusmn 1021 24278 plusmn 1718
Values are means plusmn SD of 5 animals per group HC cholesterol at 30mg03mLAA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkglowastSignificantly different from control (119875 lt 005)lowastlowastSignificantly different from HC (119875 lt 005)
Table 7 Changes in antiatherogenic coronary risk and cardioprotective indices of hypercholesterolemic rats treated with methanol extractof Artocarpus altilis and Questran for nine weeks
Treatment SerumAAI () CRI CPI
Control 2921 plusmn 212 134 plusmn 003 115 plusmn 005
HC 280 plusmn 37lowast
453 plusmn 025lowast
040 plusmn 006lowast
HC + AA1 272 plusmn 27lowast
465 plusmn 043lowast
030 plusmn 005lowast
HC + AA2 1161 plusmn 173lowastlowast
186 plusmn 017lowastlowast
089 plusmn 015lowastlowast
HC + QUE 1430 plusmn 218lowastlowast
170 plusmn 022lowastlowast
072 plusmn 008lowastlowast
QUE 1491 plusmn 121 167 plusmn 026 083 plusmn 010
AA2 2330 plusmn 227 143 plusmn 022 106 plusmn 041
HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkgAA2 Artocarpus altilis at 200mgkg QUE questran at 026 gkgantiatherogenic index (AAI) 100 times [HDL-Ctotal cholesterolminusHDL-C]coronary risk index (CRI) total cholesterolHDL-Ccardioprotective index (CPI) HDL-CLDL-ClowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005)
atherogenic process therapeutic intervention to raise HDL-C together with other risk factors is widely encouraged Inthis study treatment with AA led to significant elevationof HDL-C indicating its promising protective role againstCVD The protective roles of HDL-C from CVD have beensuggested to occur in various ways [45] HDL exerts partof its antiatherogenic effect by counteracting LDL oxidationand studies also showed that HDL promotes the reverse
cholesterol transport pathway by inducing an efflux of excessaccumulated cellular cholesterol and prevents the generationof an oxidatively modified LDL [46] Furthermore HDL notonly inhibits the oxidation of LDLby transitionmetal ions butalso prevents 12-lipoxygenase-mediated formation of lipidhydroperoxides [45] On the basis of our results AA mayprobably plays an antiatherogenic role through the inhibitionof lipids oxidation due to its antilipoperoxidative effect
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
Advances in Pharmacological Sciences 7
0
50
100
150
200
250
300
350
400C
onc
(mg
dL)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver total cholLiver trig
lowast
lowast
lowast
lowast
lowastlowastlowastlowast lowastlowast
lowastlowast
Figure 2 Effects of methanol extract of Artocarpus artilis andQuestran on hepatic total cholesterol and triglyceride levels ofhypercholesterolemic rats lowastSignificantly different from control (119875 lt005) lowastlowastsignificantly different from HC (119875 lt 005) HC hyper-cholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
observed in this study aswell as the elevation ofHDL-C LDL-C another primary target of CVD risk reduction therapy [41]In this study AA administered at a dose of 200mgkg loweredLDL-C levels of hypercholesterolemic rats It is known thatexcess of LDL can be deposited on the blood vessel wallsand becomes a major component of atherosclerotic plaquelesions Therefore serum LDL-C level has been used tomonitor treatment of patients with elevated blood cholesterollevels In view of our results AA elicited beneficial effectsby lowering serum total cholesterol including low-densitylipoprotein of the hypercholesterolemic rats In additionhypocholesterolemic and hypotriglyceridemic effects of AAmay probably be due to the inhibition of rate-limiting enzyme3-hydroxy-3-methyl glutaryl CoA reductase (HMG-CoAreductase) of cholesterol biosynthesis The experimentallyobtained hypotriglyceridemic effect of AA may also be dueto the improvement in lipolysis by reducing the activity ofhormone-sensitive lipase [42] To further support the lipidlowering potential of AA the antiatherogenic index (AAI)was also evaluated and found to increase in HC rats treatedwith AA as compared to controls Similarly improvementwas also observed in cardioprotective index (CPI) of HC ratstreated with AA in terms of HDL-CLDL-C ratio relative tocontrols Out of the risk indices considered HDL-CLDL-C ratio (CPI) was found ideal in the present study It hasbeen reported that a decrease in HDL-CLDL-C ratio is
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
lowast
lowast
lowastlowast
lowastlowast
Figure 3 Effects of methanol extract of Artocarpus artilis on theactivities of serum lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
0
10
20
30
40
50
60
70
80
LDH
(UL
)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2lowast
lowast
lowastlowast
lowastlowast
Figure 4 Effects of methanol extract of Artocarpus artilis on theactivities of cardiac lactate dehydrogenase (LDH) of hypercholes-terolemic rats lowastSignificantly different from control (119875 lt 005)lowastlowastsignificantly different from HC (119875 lt 005) HC hypercholes-terolemic rats AA1Artocarpus altilis at 100mgkg AA2Artocarpusaltilis at 200mgkg and QUE Questran at 026 gkg
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
8 Advances in Pharmacological Sciences
0
0005
001
0015
002
0025
003
0035
004
0045
GST
(120583m
ole
min
mg
prot
ein)
Treatments
Con
trol
HC
HC
+ A
A1
HC
+ A
A2
HC
+ Q
UE
QU
E
AA
2
Liver GSTHeart GST
lowast
lowastlowast
lowastlowast
lowastlowast
Figure 5 Effects of methanol extract of Artocarpus artilis on the activities of hepatic and cardiac glutathione-s-transferase (GST)of hypercholesterolemic rats lowastSignificantly different from control (119875 lt 005) lowastlowastsignificantly different from HC (119875 lt 005) HChypercholesterolemic rats AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUE Questran at 026 gkg
Control HC HC + AA1 HC + AA2
AA2QUE + AA2 QUE
Figure 6 Changes in histology of liver samples of hypercholesterolemic rats treated withArtocarpus altilis andQuestran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrow shows portal congestion periportal cellular infiltration and vacuolar degeneration of hepatocytes
good predictor of CVD in subjects [47] Similarly coronaryrisk index (CRI) in terms of TCHDL-C ratio significantlydecreased in AA-treated HC rats which further strengthenthe beneficial effects of AA
Serum AST and ALT are the reliable markers for liverfunction while serumLDHmay give information on the stateof the cardiac tissue It is established that AST can be found
in the liver cardiac muscle skeletal muscle and so forthwhereas ALT is predominantly present in the liver [48] Theincreased levels of serum AST and ALT in HC rats indicatean increased permeability and damage andor necrosis ofhepatocytes Similar results were reported by Suk et al [49]and Mohd Esa et al [50] in which ALT and AST activitieswere elevated in HC rats In our study we found that extract
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
Advances in Pharmacological Sciences 9
Control HC HC + AA1 HC + AA2
HC + QUE QUE AA2
Figure 7 Changes in histology of aorta from hypercholesterolemic rats treated with Artocarpus altilis and Questran for nine consecutiveweeks (M times400) HC cholesterol at 30mg03mL AA1 Artocarpus altilis at 100mgkg AA2 Artocarpus altilis at 200mgkg and QUEQuestran at 026 gkg Black arrows shows myofibril necrosis severe hemorrhages and fibrous connective tissue laid down
of AA at a dose of 200mgkg caused a significant reductionin the activities of serum AST ALT and LDH which furthersupports the beneficial effects of the extract of AA in HC rats
Oxidative stress defined as a disruption of the balancebetween oxidative and antioxidative processes plays animportant role in the pathogenesis of atherosclerosis [51]Studies in animal models and human clinical trials haveestablished a relationship between hypercholesterolemia andlipid peroxidation [50] In agreement with these findings ourresults show increased levels ofMDA in the serum and tissuesof HC rats when compared to controls On the other handtreatment with AA caused a significant reduction in the levelsof MDA in these organs This protective effect is probablybased on the antioxidant activity of AA which reduced theoxidative damage by blocking the production of free radicalsand thus inhibited lipid peroxidation In this study we alsoobserved a significant decrease in the activities of free radicalscavenging enzymes SOD andCAT which are the first line ofdefence against oxidative injuryThe inhibition of antioxidantsystem (SOD and CAT)may cause the accumulation of H
2O2
or products of its decomposition [52] SOD catalyzes theconversion of superoxide anion into H
2O2 The primary role
of CAT is to scavenge H2O2that has been generated by
free radical or by SOD Importantly administration of AArestored the activities of enzymatic antioxidants (SOD andCAT) in liver and heart of HC rats AA may therefore act asan effective antioxidant of great importance against diseasesand degenerative processes caused by oxidative stress Ourresults showed that the extract of AA at 750 120583gmL produced63 inhibition of DPPH radical relative to catechin (68)The antioxidant property of AA may be linked to highpolyphenolic compounds in this plant as shown in our resultsFrom these findings AA positivelymodulates the antioxidantredox status of HC rats in addition to its beneficial effects onthe lipid profile
5 Conclusion
The present study suggests that Artocarpus altilis has potentblood and tissues lipid-lowering capability In additionit has significant antiatherogenic effect and also improvesantioxidant system of hypercholesterolemic rats Furtherstudies are required to identify the active component(s) andmechanism(s) underlying the beneficial effects of this plant
Conflict of Interests
The authors declare that they have no conflict of interests
Acknowledgments
This research was partly supported by Senate ResearchGrants fromUniversity of Ibadan Nigeria Oluwatosin Adek-unle given to Dr Oluwatosin Adekunle Adaramoye (SRGCOM20107A)
References
[1] P Kresanov M Ahotupa T Vasankari et al ldquoThe associationsof oxidized high-density lipoprotein lipids with risk factors foratherosclerosis the cardiovascular risk in young finns studyrdquoFree Radical Biology and Medicine C vol 65 pp 1284ndash12902013
[2] K Kovacs K Erdelyi C Hegedus et al ldquoPoly(ADP-ribo-syl)ation is a survival mechanism in cigarette smoke-inducedand hydrogen peroxide-mediated cell deathrdquo Free RadicalBiology and Medicine vol 53 no 9 pp 1680ndash1688 2012
[3] G Sikka D Pandey A K Bhuniya et al ldquoContribution ofarginase activation to vascular dysfunction in cigarette smok-ingrdquo Atherosclerosis vol 231 no 1 pp 91ndash94 2013
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
10 Advances in Pharmacological Sciences
[4] M Fuzi Z Palicz J Vincze et al ldquoFluvastatin-induced alter-ations of skeletal muscle function in hypercholesterolaemicratsrdquo Journal of Muscle Research and Cell Motility vol 32 no6 pp 391ndash401 2012
[5] N A Salem and E A Salem ldquoRenoprotective effect of grapeseed extract against oxidative stress induced by gentamicin andhypercholesterolemia in ratsrdquo Renal Failure vol 33 no 8 pp824ndash832 2011
[6] T Rantanen ldquoMidlife fitness predicts less burden of chronicdisease in later liferdquo Clinical Journal of Sport Medicine vol 23no 6 pp 499ndash500 2013
[7] J A Borke and P C Wyer ldquoEating a larger number of high-salt foods is not associated with short-term risk of acutedecompensation in patients with chronic heart failurerdquo TheJournal of Emergency Medicine vol 44 no 1 pp 36ndash45 2013
[8] R Schekman ldquoDiscovery of the cellular and molecular basisof cholesterol controlrdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 37 pp14833ndash14836 2013
[9] C AMiller ldquoUpdate on statins and other lipid-lowering drugsrdquoGeriatric Nursing vol 22 no 5 pp 276ndash277 2001
[10] AM Rincon L B Rached L E Aragoza and F Padilla ldquoEffectof acetylation and oxidation on some properties of Breadfruit(Artocarpus altilis) seed starchrdquo Archivos Latinoamericanos deNutricion vol 57 no 3 pp 287ndash294 2007
[11] N J C Zerega D Ragone and T J Motley ldquoSystematics andspecies limits of breadfruit (Artocarpus moraceae)rdquo SystematicBotany vol 30 no 3 pp 603ndash615 2005
[12] C A Lans ldquoEthnomedicines used in Trinidad and Tobago forurinary problems and diabetes mellitusrdquo Journal of Ethnobiol-ogy and Ethnomedicine vol 2 pp 45ndash56 2006
[13] C R Nwokocha D U OwuMMcLaren et al ldquoPossiblemech-anisms of action of the aqueous extract of Artocarpus altilis(breadfruit) leaves in producing hypotension in normotensiveSprague-Dawley ratsrdquo Pharmaceutical Biology vol 50 no 9 pp1096ndash1102 2012
[14] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 no 1 pp 152ndash178 1998
[15] L I Mensor F S Menezes G G Leitao et al ldquoScreening ofBrazilian plant extracts for antioxidant activity by the use ofDPPH free radical methodrdquo Phytotherapy Research vol 15 no2 pp 127ndash130 2001
[16] O A Adaramoye VONwaneri K C Anyanwo EO Farombiand G O Emerole ldquoPossible anti-atherogenic effect of kolav-iron (a Garcinia kola seed extract) in hypercholesterolaemicratsrdquo Clinical and Experimental Pharmacology and Physiologyvol 32 no 1-2 pp 40ndash46 2005
[17] O H Lowry N J Rosebrough A L Farr and R J RandallldquoProtein measurement with the Folin phenol reagentrdquo TheJournal of Biological Chemistry vol 193 no 1 pp 265ndash275 1951
[18] A F Mohun and I J Cook ldquoSimple methods for measuringserum levels of the glutamic-oxalacetic and glutamic-pyruvictransaminases in routine laboratoriesrdquo Journal of ClinicalPathology vol 10 no 4 pp 394ndash399 1957
[19] S Reitman and S Frankel ldquoA colorimetricmethod for the deter-mination of serum glutamic oxalacetic and glutamic pyruvictransaminasesrdquo American Journal of Clinical Pathology vol 28no 1 pp 56ndash63 1957
[20] W Richmond ldquoPreparation and properties of a cholesteroloxidase from Nocardia sp and its application to the enzymaticassay of total cholesterol in serumrdquo Clinical Chemistry vol 19no 12 pp 1350ndash1356 1973
[21] N J Jacobs and P J van Demark ldquoThe purification and proper-ties of the 120572-glycerophosphate-oxidizing enzyme of Streptococ-cus faecalis 10C1rdquo Archives of Biochemistry and Biophysics vol88 no 2 pp 250ndash255 1960
[22] L K Koditschek and W W Umbreit ldquoAlpha-glycerophosphateoxidase in Streptococcus faecium F 24rdquo Journal of Bacteriologyvol 98 no 3 pp 1063ndash1068 1969
[23] W T Friedewald R I Levy and D S Fredrickson ldquoEstimationof the concentration of low-density lipoprotein cholesterol inplasma without use of the preparative ultracentrifugerdquo ClinicalChemistry vol 18 no 6 pp 499ndash502 1972
[24] J A Buege and S D Aust ldquoMicrosomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978
[25] H J Zimmerman and H G Weinstein ldquoLactic dehydrogenaseactivity in human serumrdquoThe Journal of Laboratory andClinicalMedicine vol 48 no 2 pp 607ndash609 1956
[26] J M McCord and I Fridovich ldquoSuperoxide dismutase Anenzymic function for erythrocuprein (hemocuprein)rdquoThe Jour-nal of Biological Chemistry vol 244 no 22 pp 6049ndash6055 1969
[27] H Aebi ldquoCatalase estimationrdquo in Methods of Enzymatic Anal-ysis H V Bergmeyer Ed pp 673ndash684 Verlag Chemic NewYork NY USA 1974
[28] E Beutler O Duron and B M Kellin ldquoImproved methodfor the determination of blood glutathionerdquo The Journal ofLaboratory and Clinical Medicine vol 61 pp 882ndash888 1963
[29] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973
[30] W H Habig M J Pabst and W B Jakoby ldquoGlutathioneS transferases The first enzymatic step in mercapturic acidformationrdquoThe Journal of Biological Chemistry vol 249 no 22pp 7130ndash7139 1974
[31] P Barter AM Gotto J C LaRosa et al ldquoHDL cholesterol verylow levels of LDL cholesterol and cardiovascular eventsrdquo TheNew England Journal of Medicine vol 357 no 13 pp 1301ndash13102007
[32] H-T Kang J-K Kim J-Y Kim J A Linton J-H Yoon and S-B Koh ldquoIndependent association of TGHDL-C with urinaryalbumin excretion in normotensive subjects in a rural Koreanpopulationrdquo Clinica Chimica Acta vol 413 no 1-2 pp 319ndash3242012
[33] A A Adeneye and J A Olagunju ldquoPreliminary hypoglycaemicand hypolipidemic activities of the aqueous seed extract ofCarica papaya Linn in Wistar ratsrdquo Biology and Medicine vol1 no 1 pp 1ndash10 2009
[34] M A Waqar and Y Mahmmod ldquoAnti-platelet anti-hypercholesterolemia and anti-oxidant effects of Ethanolicextract of Brassica oleracea in high fat diet provided ratsrdquoWorldApplied Sciences Journal vol 8 no 1 pp 107ndash112 2010
[35] S Paolillo G L Della Ratta A Vitagliano et al ldquoNewperspectives in cardiovascular risk reduction focus on HDLrdquoArchives for Chest Disease vol 80 no 1 pp 27ndash30 2013
[36] S C Smith Jr E J Benjamin R O Bonow et al ldquoAHAACCFsecondary prevention and risk reduction therapy for patientswith coronary and other atherosclerotic vascular diseaserdquoCirculation vol 124 no 22 pp 2458ndash2473 2011
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
Advances in Pharmacological Sciences 11
[37] Z Reiner A L Catapano G de Backer et al ldquoESCEASGuidelines for the management of dyslipidaemiasrdquo EuropeanHeart Journal vol 32 no 14 pp 1769ndash1818 2011
[38] K Yuji H Sakaida T Kai et al ldquoEffect of dietary blueberry(VacciniumasheiReade) leaves on serumandhepatic lipid levelsin ratsrdquo Journal of Oleo Science vol 62 no 2 pp 89ndash96 2013
[39] O A Adaramoye O Akintayo J Achem and M A FafunsoldquoLipid-lowering effects of methanolic extract ofVernonia amyg-dalina leaves in rats fed on high cholesterol dietrdquo VascularHealth and Risk Management vol 4 no 1 pp 235ndash241 2008
[40] V Kamesh and T Sumathi ldquoAntihypercholesterolemic effectof Bacopa monniera linn on high cholesterol diet inducedhypercholesterolemia in ratsrdquo Asian Pacific Journal of TropicalMedicine vol 5 no 12 pp 949ndash955 2012
[41] P O Kwiterovich Jr ldquoThe metabolic pathways of high-densitylipoprotein low-density lipoprotein and triglycerides a currentreviewrdquoAmerican Journal of Cardiology vol 86 no 12 pp 5ndash102000
[42] M L Bishop E P Fody and L SchoeffClinical Chemistry Prin-ciples Procedures Correlations Lippincott Williams ampWilkins6th edition 2010
[43] S A Qureshi M Kamran M Asad A Zia T Lateef andM BAzmi ldquoA preliminary study of Santalum album on serum lipidsand enzymesrdquo Global Journal of Pharmacology vol 4 no 2 pp71ndash74 2010
[44] T D Filippatos and M S Elisaf ldquoHigh density lipoprotein andcardiovascular diseasesrdquoWorld Journal of Cardiology vol 5 no7 pp 210ndash214 2013
[45] J-R Nofer B Kehrel M Fobker B Levkau G Assmann andA V Eckardstein ldquoHDL and arteriosclerosis beyond reversecholesterol transportrdquo Atherosclerosis vol 161 no 1 pp 1ndash162002
[46] T Yokozawa E J Cho S Sasaki A Satoh T Okamoto and YSei ldquoThe protective role of Chinese prescription Kangen-karyuextract on diet-induced hypercholesterolemia in ratsrdquoBiologicaland Pharmaceutical Bulletin vol 29 no 4 pp 760ndash765 2006
[47] T Marotta B F Russo and L A Ferrara ldquoTriglyceride-to-HDL-cholesterol ratio and metabolic syndrome as contributorsto cardiovascular risk in overweight patientsrdquo Obesity vol 18no 8 pp 1608ndash1613 2010
[48] R Rej ldquoLiver diseases and the clinical laboratorymdashthe twen-tieth Arnold O Beckman conference in clinical chemistryrdquoClinical Chemistry vol 43 no 8 pp 1473ndash1475 1997
[49] F-M Suk S-Y Lin C-H Chen et al ldquoTaiwanofungus cam-phoratus activates peroxisome proliferator-activated receptorsand induces hypotriglyceride in hypercholesterolemic ratsrdquoBioscience Biotechnology and Biochemistry vol 72 no 7 pp1704ndash1713 2008
[50] N Mohd Esa K K Abdul Kadir Z Amom and A AzlanldquoAntioxidant activity of white rice brown rice and germinatedbrown rice (in vivo and in vitro) and the effects on lipidperoxidation and liver enzymes in hyperlipidaemic rabbitsrdquoFood Chemistry vol 141 no 2 pp 1306ndash1312 2013
[51] A V Rzheshevsky ldquoFatal ldquotriadrdquo lipotoxicity oxidative stressand phenoptosisrdquo Biochemistry vol 78 no 9 pp 991ndash10002013
[52] C E Cross A van der Vliet C A OrsquoNeill S Louie and BHalliwell ldquoOxidants antioxidants and respiratory tract liningfluidsrdquo Environmental Health Perspectives vol 102 no 10 pp185ndash191 1994
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom
Volume 2014
ToxinsJournal of
VaccinesJournal of
Hindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AntibioticsInternational Journal of
ToxicologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Drug DeliveryJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in Pharmacological Sciences
Tropical MedicineJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Medicinal ChemistryInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
AddictionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Autoimmune Diseases
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anesthesiology Research and Practice
ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Pharmaceutics
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
MEDIATORSINFLAMMATION
of