The Journal of Basic & Applied Zoology (2015) 72, 154–162

HO ST E D BYThe Egyptian German Society for Zoology

The Journal of Basic & Applied Zoology

www.egsz.orgwww.sciencedirect.com

Renoprotective effect of Mangifera indicapolysaccharides and silymarin against

cyclophosphamide toxicity in rats

* Corresponding author.

Peer review under responsibility of The Egyptian German Society for

Zoology.

http://dx.doi.org/10.1016/j.jobaz.2015.09.0062090-9896 � 2015 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V.This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Ahmed I. Amien a, Sohair R. Fahmy b, Fathi M. Abd-Elgleel c,

Sara M. Elaskalany b,*

aDepartment of Biochemistry, Faculty of Science, Cairo University, EgyptbDepartment of Zoology, Faculty of Science, Cairo University, EgyptcDepartment of Chemistry, Faculty of Science, Cairo University, Egypt

Received 3 May 2015; revised 28 September 2015; accepted 30 September 2015Available online 10 December 2015

KEYWORDS

Mangifera indica L.;

Polysaccharides;

Oxidative stress;

Cyclophosphamide;

Silymarin;

Nephrotoxicity

Abstract The present study aims to evaluate the possible protective role of polysaccharides

extracted from the Egyptian mango Mangifera indica L. (MPS) and silymarin against cyclophos-

phamide (CP) nephrotoxicity in male albino rats. Male rats were randomly divided into, control

group (administered distilled water orally for 10 days) and MPS (500, 1000 mg/kg, p.o.) and/or sily-

marin (150 mg/kg, p.o.) treated groups for 10 days. In the last 5 days of treatment rats were admin-

istered CP (150 mg/kg, i.p). The MPS revealed significant prophylactic effect against kidney injury

induced by CP as demonstrated by enhancement of the kidney function via decreasing serum cre-

atinine, urea and uric acid. Treatment of rats with MPS extract and/or silymarin significantly

increased the level of reduced glutathione (GSH) and superoxide dismutase (SOD) activity while

decreased the level of total malondialdehyde (MDA) and glutathione-S-transferase (GST). Also,

histopathological examinations confirmed the protective efficacy of MPS and/or silymarin against

CP nephrotoxicity. In conclusion, the obtained results of the present study support the protective

antioxidant role of MPS and/or silymarin against CP-induced kidney disorder in rats.� 2015 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V. This is an

open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

The kidney is a primary target for numerous toxic xenobioticsincluding drugs, environmental chemicals and metals. Acute

kidney injury, induced by drugs and other stimuli, is a signifi-

cant clinical problem, and accounts for the cessation of devel-opment of many promising drug candidates Shelton et al.(2013). Drug-induced nephrotoxicity is an extremely common

condition and is responsible for a variety of pathologicaleffects on the kidneys Dhodi et al. (2014). Cyclophosphamide(CP) is a potent anticancer agent. CP is effective against a widespectrum of malignancies, such as leukemia, lymphoma,

breast, lung, prostate, and ovarian cancers Khan et al.(2004). The usage of CP is severely limited by its physiological

Renoprotective effect of Mangifera indica polysaccharides and silymarin 155

side effects, such as hepatotoxicity, nephrotoxicity, urotoxic-ity, cardiotoxicity and myelosuppression Motawi et al.(2010), Lameire et al. (2011) and Newton (2012). However,

its clinical use is restricted because of its marked organ toxicityassociated with increased oxidative stress and inflammationNafees et al. (2015). Oxidative stress is reported to play impor-

tant roles in CP-induced renal damage Abraham and Rabi(2011). Renal damage is one of the dose-limiting side effectsof CP Abraham and Rabi (2011). It has been demonstrated

that increased generation of reactive oxygen species (ROS)by CP in kidney tissues plays a critical role in the pathogenesisof CP-induced kidney damage Stankiewicz and Skrzydlewska(2003) and Abraham and Rabi (2009). So, free radical scav-

engers and antioxidants can be used in the treatment Sabolic(2006). Cyclophosphamide is now being used in combinationwith various detoxifying and protective agents with the pur-

pose of reducing or eliminating its adverse toxic effectsNeboh and Ufelle (2015) and Nurrochmad et al. (2015).

Silymarin, a bioflavonoid, is the main constituent of Sily-

bummarianum (milk thistle). Chemically, silymarin is aflavonolignan that consists of a mixture of mainly three flavo-noids, silibinin, silydianin and silychristin Kiruthiga et al.

(2007). Silymarin has been reported to possess several pharma-cological activities including antioxidant and anti-inflammatory/immunomodulatory Manna et al. (1999), antifibroticCrocenzi and Roma (2006), hepatoprotective, antibacterial,

antiallergic, antimutagenic, antiviral, antithrombotic andvasodilatory actions Wellington and Jarvis (2001). Many stud-ies reported that silymarin reduced CP-induced ROS genera-

tion Jnaneshwari et al. (2012). Silymarin was shown to havepositive effects on preventing or decreasing severity ofcyclophosphamide nephrotoxicity Eser et al. (2012). So it is

the positive control used in this study.Pharmacotherapy using natural substances can be currently

regarded as a very promising future alternative to conventional

therapyWang et al. (2013). Plants have been themajor source oftherapeutic agents for curing the human diseases Zahid et al.(2013). Mango (Mangifera indica L.) is one of the most impor-tant and popular tropical fruits, mainly due to its attractive fla-

vor Chen et al. (2012). M. indica L. is an important member ofthe family Anacardiaceae and belongs to genusMangifera ordersapindales Ross (1999). In recent years, some bioactive polysac-

charides isolated from natural sources have attracted muchattention in the field of biochemistry and pharmacology Wanget al. (2013) andXue et al. (2015). Some polysaccharides isolated

from natural sources show various important biologicalactivities, such as antitumor, immunomodulatory, andanti-inflammatory effects, which are strongly affected by theirchemical structures and chain conformations Liu et al. (2015).

It has been well documented that polysaccharides increaseimmunity through production of interleukins and antibodiesYang et al. (2008) and therefore could be explored as novel

prospective antioxidants Sun et al. (2010) and Chen et al.(2012). Polysaccharides can be currently regarded as a verypromising future alternative to conventional therapy in kidney

diseases Chiu et al. (2014) and Lu et al. (2014). When multipleantioxidants are used in combination, they protect againstvulnerability to other agents and synergistically potentiate their

antioxidant properties Aleisa et al. (2013). So, the present studywas designed to evaluate the efficacy of polysaccharidesextracted from the mango pulp (MPS) and/or silymarin againstrenal injury induced by cyclophosphamide in male albino rats.

Materials and methods

Chemicals and reagents

Methanol 95%, ethanol, absolute acetone, and cyclophos-phamide (Endoxan) were purchased from Bexter Oncology

GmbH (Germany), silymarin was purchased from SEDICOPharmaceutical Co. (6 October City, Egypt). All other chemi-cals were of analytical grade. Kits for all biochemical parame-

ters were purchased from Biodiagnostic Company (El-Dokki,Giza, Egypt).

Collection of Mangifera indica L.

Mature green mango (M. indica L.) cv. Fagrkelan fruits werefreshly harvested from Egypt fields through a local Dealer inSeptember 2012, M. indica washed thoroughly with running

tap water, seeds and peels are removed from mango.

Extraction of polysaccharides from Mangifera indica L.

Polysaccharides extracted from mango pulp by the techniquedescribed by Devaki et al. (2009) with some modifications.Briefly, 1670 g of mango pulp was boiled in 16.7 L of distilled

water at 100 �C for 3 h. The slurry was separated by gauze andthen, was filtered. The filtrate was dialyzed against tap waterfor 48 h, and then concentrated to about 350 ml under reducedpressure using rotary evaporator. Then, the concentrated fil-

trate was precipitated using four volumes of 95% ethanol toone volume of the extract (about 1400 ml 95% ethanol). Theextract was washed with absolute acetone, filtrated and then

dried in a vacuum desiccator over anhydrous calcium chloride.About 65 g of polysaccharides extract is obtained and used foranimals’ experiments.

Animals

The experimental animals used in this study were adult male

albino Wistar rats (Rattus norvegicus) weighing (150–160± 5 g). The animals were obtained from the NationalResearch Center (NRC, Dokki, Giza). Animals were groupedand housed in polyacrylic cages (Five animals per cage). Ani-

mals were given food and water ad libitum. Rats were main-tained in a friendly environment with a 12 h/12 h light–darkcycle at room temperature (22–25 �C). Rats were acclimatized

to laboratory conditions for 7 days before commencement ofthe experiment. The protocol was approved by the CairoUniversity, Faculty of Science Institutional Animal Care and

Use Committee (IACUC), Egypt (CUFS/S/PHY/28/14), andall the experimental procedures were carried out in accordancewith international guidelines for care and use of laboratoryanimals.

Toxicity study (OECD 420)

Adult male albino rats (R. norvegicus) weighing (150–160

± 5 g) were used for acute toxicity studies. The rats weredivided into control and test groups containing five animalseach. The rats were administered orally with polysaccharides

extract of mango at dose levels of 5000 mg/kg (high dose)

156 A.I. Amien et al.

and 2000 mg/kg (low dose). Normal control rats received thesame amount of vehicle (distilled water) only. Rats wereobserved carefully for 24 h after extract administration and

then for the next 14 days. At the end of this experimental per-iod, the rats were observed for signs of toxicity, morphologicalbehavior, and mortality. Acute toxicity was evaluated based

on the number of deaths (if any). Acute toxicity was calculatedas per Organization for Economic Co-operation and Develop-ment (OECD) guidelines 420 (Fixed dose method) (Van den

Heuvel et al., 1990; Whitehead and Curnow, 1992).

Experimental design

Thirty-five rats were randomly divided into seven groups (5 ani-mals/group) as follows: Group 1: Served as a control group.Group 2: Rats were administered distilled water (2 ml) orallyfor 10 days. Group 3: Rats treated orally with MPS (dissolved

in distilled water) in a dosage of 500 mg/kg body weight for10 days. Group 4: Rats treated orally with MPS in a dosage of1000 mg/kg body weight for 10 days. Group 5: Rats treated

orally with silymarin (dissolved in distilled water) in a dosageof 150 mg/kg body weight for 10 days. Group 6: Rats treatedorally with MPS in a dosage of 500 mg/kg body weight and

silymarin 150 mg/kg body weight at the same time for10 days. Group 7: Rats treated orally with MPS in a dosage of1000 mg/kg body weight and silymarin 150 mg/kg body weightat the same time for 10 days (see Fig. 1).Rats of all groups except

group one treated intraperitoneally with CP in a dosage of150 mg/kg body weight in the last 5 days (Jnaneshwari et al.,2012).

Experimental procedures

After 10 days of treatment, the animals were fasted overnight

(12–14 h). Rats of each group were sacrificed by exsanguina-tions under sodium pentobarbital anesthesia, and the bloodsamples were collected from each animal into sterilized cen-

trifuge tubes for serum separation. Kidney was removed andimmediately blotted using filter paper to remove traces ofblood. The first part was immersed in 10% formal saline solu-tion for fixation preparatory histopathological examinations.

The other part of kidney was stored at �80 �C for biochemicalanalysis. Blood samples were centrifuged at 3000 rpm for15 min. Serum was separated and stored in Eppendorf at

�20 �C to be used for biochemical analysis. The kidney washomogenized (10% w/v) in ice cold 0.1 M phosphate buffer(pH 7.4). The homogenate was centrifuged at 3000 rpm for

15 min at 4 �C and the resultant supernatant was used for esti-mation of oxidative stress markers.

Assessment of serum biochemical parameter

All kits for biochemical analysis were purchased from Biodiag-nostic Company. Determination of serum creatinine accordingto colorimetric method was described by Tietz and Andresen

(1986). Urea was determined in serum by modified urease-Berthlot method according to Tietz et al. (1990). Serum uricacid was determined colorimetrically according to Tietz (1990).

Assessment of oxidative stress markers

Oxidative stress markers were detected in the resultant super-natant of kidney homogenate by using Biodiagnostic kits.Lipid peroxide (Malondialdehyde) MDA was determined in

tissue according to Ohkawa et al. (1979). Reduced glutathione(GSH) was determined in tissue according to Beutler et al.(1963). Superoxide dismutase (SOD) activity was determinedaccording to Nishikimi et al. (1972). Glutathione-S-

transferase (GST) activity was determined according toHabig et al. (1974).

Histopathological analysis

Histological sections (4 lm thick) were prepared from paraffinblocks of kidney tissues fixed in 10% formal saline. Sections

were stained with hematoxylin and eosin (H&E) (Mayer,1903) and then, examined under light microscope under�400 magnifications.

Statistical analysis

Values were expressed as mean ± SEM. (standard error) toevaluate differences between the groups studied, a one way

analysis of variance (ANOVA) with the Duncan post hoc testwas used to compare the group means and P < 0.05 was con-sidered statistically significant. SPSS for Windows (version

19.0) was used for the statistical analysis.

Results

Toxicity study

Nevertheless, none of the five rats of the test group died anddid not show any sign of toxicity after administration ofMPS at higher dosage (5000 mg/kg) in the first 24 h or during

the experiment period (14 day) and the activities of serumALAT and serum ASAT are at normal range compared withthe control group. In addition, there is no histopathologicalchange in both liver and kidney. The single lethal dose of

MPS that kills half of the animals (LD50) was therefore takenas above 2000 mg/kg. MPS is tested at two dosage levels (500and 1000 mg/kg) as they represented 1/10th and 1/5th of the

proposed LD50.

Improvement of kidney function in M. indica polysaccharides(MPS) extract and/or silymarin treated rats

As shown in (Table 1), serum creatinine, urea and uric acidlevels were significantly increased (P < 0.05) following admin-

istration of CP compared to the control group. However,treatment with MPS (500, 1000 mg/kg) and/or silymarin signif-icantly showed enhancement in the recorded kidney functionmarkers.

Figure 1 Schematic diagram shows the experimental design of treatment.

Table 1 Effect of Mangifera indica polysaccharides (MPS) extract and/or silymarin on kidney functions parameter in serum of

cyclophosphamide (CP) – treated rats.

Groups Serum creatinine (mg/dL) Serum urea (mg/dL) Serum uric acid (mg/dL)

Control 57 ± 2.54a 123.34 ± 5.03bc 3.442 ± 0.108a

CP Vehicle(saline) 81 ± 1.87d 219.18 ± 11.74d 4.406 ± 0.135b

MPS (500 mg/kg) 69.2 ± 1.41c 94.16 ± 5.37a 3.3 ± 0.133a

MPS (1000 mg/kg) 71.5 ± 1.87c 102.52 ± 1.02ab 3.328 ± 0.058a

Silymarin 63.5 ± 4.71abc 127.5 ± 13.42c 3.19 ± 0.081a

MPS (500 mg/kg)silymarin 68 ± 2.427bc 104.16 ± 5.43abc 3.438 ± 0.132a

MPS (1000 mg/kg) + silymarin 60.7 ± 1.72ab 113.34 ± 3.58abc 3.272 ± 0.086a

Values are given as mean ± SEM for 5 rats in each group. Values with different superscript letters are significantly different at P < 0.05.

Figure 2 Effect of Mangifera indica polysaccharides (MPS)

extract and/or silymarin on kidney malondialdehyde (MDA) in

serum of cyclophosphamide (CP) – treated rats. Values are given

as mean ± SEM for 5 rats in each group. Values with different

superscript letters are significantly different at (P < 0.05).

Figure 3 Effect of Mangifera indica polysaccharides (MPS)

extract and/or silymarin on kidney glutathione reduced (GSH) in

serum of cyclophosphamide (CP) – treated rats. Values are given

as mean ± SEM for 5 rats in each group. Values with different

superscript letters are significantly different at (P < 0.05).

Renoprotective effect of Mangifera indica polysaccharides and silymarin 157

Figure 5 Effect of Mangiferaindica polysaccharides (MPS)

extract and/or silymarin on kidney glutathione-S-transferase

(GST) in serum of cyclophosphamide (CP) – treated rats. Values

are given as mean ± SEM for 5 rats in each group. Values with

different superscript letters are significantly different at

(P< 0.05).

Figure 4 Effect of Mangifera indica polysaccharides (MPS)

extract and/or silymarin on kidney superoxide dismutase (SOD) in

serum of cyclophosphamide (CP) – treated rats. Values are given

as mean ± SEM for 5 rats in each group. Values with different

superscript letters are significantly different at (P < 0.05).

158 A.I. Amien et al.

Improvement of some oxidative parameters in M. indicapolysaccharides (MPS) extract and/or silymarin treated rats

Administration of CP significantly (P < 0.05) increased the

level of malondialdehyde (MDA) and the activity ofglutathione-S-transferase (GST) in the kidney tissue comparedto the control group (Figs. 2 and 5). On the other hand, CP

caused significant decrease (P < 0.05) in reduced glutathione(GSH) level and superoxide dismutase (SOD) activity com-pared to the control group (Figs. 3 and 4). Treatment of ratseither with MPS extract (500 and 1000 mg/kg body weight)

and/or silymarin significantly increased (P < 0.05) the levelof GSH level. Level of MDA and activity of GST significantlydecreased following treatment of rats either with MPS extract

(500 and 1000 mg/kg body weight) and/or silymarin (Figs. 2and 5). Administration of MPS extracts (500 mg/kg bodyweight) and/or silymarin significantly increased the activities

of SOD (Fig. 4). On the other hand treatment with MPSextracts (1000 mg/kg body weight) and/or silymarin failed toincrease the SOD activity (Fig. 4).

Kidney histopathological analysis

Representative views of kidney sections are shown in (Fig. 6).

As shown in tissue sections stained with hematoxylin andeosin, CP caused dilation and congestion of renal blood vessel,vacuolations of epithelial lining renal tubules and atrophy ofglomerular tuft (Fig. 6B–D). It can also be noticed protein cast

in the lumen of renal tubules. Oral administrations of MPSextract (500 and 1000 mg/kg body weight) and/or silymarinimproved the kidneys architecture compared to the CP group

(Fig. 6E–H).

Discussion

Chemotherapeutic drugs cause renal injury at different levelsand their effect ranges from an elevation in the level of serumcreatinine to renal failure. Among which renal toxicity is a

dreadful complication developed in cancer patients uponcyclophosphamide (CP) therapy Sinanoglu et al. (2012) andRehman et al. (2012). For the therapeutic strategies of renalinjury and disease during CP treatment, it is important to find

complementary antioxidant compound that is able to blockrenal injuries. Polysaccharides chemoprevention activityagainst cyclophosphamide has been indicated in several

reports Huang et al. (2013), Zhang et al. (2013), Yu et al.(2014). The present study throws light for the first time onthe therapeutic effect of M. indica polysaccharides (MPS)

and/or silymarin against nephrotoxicity induced by CP in rats.During the progression of the renal disease, loss of kidney

function is accompanied by failing organ function leading to

accumulation of a series of compounds Vanholder et al.(2008). Hepatorenal syndrome is most frequently first diag-nosed by a finding of increasing concentrations of serum crea-tinine or blood urea nitrogen (BUN) Gine‘ s et al. (2003). The

marked increase in the levels of serum urea and creatinine is amarker for the nephrotoxicity and kidney damage Cagler et al.(2002). In consistent with the finding of Cagler et al. (2002) and

Rehman et al. (2012), the results of the present investigationshowed significant increase in the serum urea and creatininefollowing CP administration. Creatinine is an endogenous sub-

stance produced by muscle from creatine and creatine phos-phate, with a rate of production proportionate to the musclemass Kurniawan et al. (2013). The measurement of serum cre-

atinine proves useful in diagnosing renal failure and diseaseswhere, directly proportional relationship exists between crea-tinine levels and renal function Sharma (2011). This increasein serum creatinine may be due to the hepatic damage which

evolved into a stage with features of hepatorenal syndromeArroyo and Jimenez (2000) including reduction in creatinineclearance and low glomerular filtration rate Briglia and

Anania (2002).

Figure 6 Photomicrographs of hematoxylin-eosin stained kidney histology of male albino rats (�400 magnifications). (A) Kidney

section from control group shows normal histological structure of renal parenchyma with normal structure of glomerulus. (B) Kidney

section from CP treated group showing dilation and congestion of renal blood vessel (BV). (C) Kidney section from CP treated group

showing vacuolations of epithelial lining renal tubules and atrophy of glomerular tuft. (D) kidney section from CP treated group showing

protein cast (PC) in the lumen of renal tubules. (E),(F), (G) and (H) showing kidney section from the group treated with MPS (500 and

1000 mg/kg) and/or silymarin showing the regeneration of kidney parenchyma.

Renoprotective effect of Mangifera indica polysaccharides and silymarin 159

Uric acid, a product of purine metabolism, is degraded in

most mammals by the hepatic enzyme, urate oxidase (uricase),to allantoin, which is mainly excreted by the kidneys (65–75%)and intestines (25–35%) De Oliveira and Burini (2012). The

significant increase in uric acid following CP administration

in the present study may be due to reduction in glomerular fil-

tration rate and renal urate excretion Vaziri et al. (1995).Hyperuricemia also may result from increased net tubularabsorption. After filtration, uric acid undergoes both reabsorp-

tion and secretion in the proximal tubule, and this process is

160 A.I. Amien et al.

mediated by a urate/anion exchanger and a voltage sensitiveurate channel Leal-Pinto et al. (1999) and Enomoto et al.(2002). In consonance with the finding of Yang et al. (2015),

the present study showed a significant decrease in the serumcreatinine, urea and uric acid following treatment with MPS(500, 1000 mg/kg) and/or silymarin that may be a contributory

self-healing mechanism restoring the kidney structure andfunction.

Oxidative stress mediates a wide range of renal impairments

Ebrahimi et al. (2013) and Park et al. (2013). Reactive oxygenspecies (ROS) could be renotoxic consequently impairing thefunctional capacity of the kidney. Some anti-cancer drugsincluding CP are known to exert their cytotoxic effects by a

free radical mediated mechanism Dumontet et al. (2001). Lipidperoxidation is a free radical induced process leading to oxida-tive deterioration of poly unsaturated fatty acid. It comes from

the reaction of free radicals with lipids and considered to be animportant feature of the cellular injury brought by free radicalattack Hoek and Pastorino (2002). Malondialdehyde (MDA)

is the one of end products of lipid peroxidation and measureof free radical generation Sundaram and Murugesan (2011).In conjunction with the reports of Ayhanci et al. (2010) and

Jnaneshwari et al. (2012), MDA level increased significantlyin the kidney tissue of CP-treated rats as a result of lipid per-oxidation and nephrotoxicity. It is observed that MPS andsilymarin provided a significant protection against CP-

induced nephrotoxicity by decreasing the level of MDA.Reduced glutathione (GSH) is a major constituent of the

detoxification pathways. The amino acid constituent cysteine

of GSH and proteins is regarded as being the first line ofdefense, and neutralizes the hydroxyl radical and plays animportant role against inflammation and oxidative stress

Circu and Aw (2011) The high electron-donating capacity ofthe SH group of cysteine provides great reducing power andis used to regulate a complex thiol-exchange system. In agree-

ment with the reports of Atessahin et al. (2005) and Ayhanciet al. (2010), the GSH level in the kidney tissues treated withCP significantly decreased which may be due to the nephrotox-icity and accumulation of free radicals. Treatment with MPS

and silymarin shows significant increase in the level of GSHwhich may be due its antioxidant effect.

Superoxide dismutase (SOD) is the first antioxidant enzyme

to deal with oxyradicals by accelerating the dismutation ofsuperoxide to hydrogen peroxide. Catalase (CAT) is a peroxi-somal hem protein that catalyzes the removal of hydrogen per-

oxide formed during the reaction catalyzed by SOD. Thus,SOD and CAT act as mutually supportive antioxidativeenzymes that provide protective defense against ROS Cohenet al. (1978). Viewed in conjunction the finding of Jayaraman

et al. (2008), the present study shows significant decrease inthe level of SOD in the kidney of CP treated rats. Theenhanced free radical concentration resulting from the oxida-

tive stress conditions can cause loss of enzymatic activitySanzgiri et al. (1997). However, treatment with MPS in thepresent study restored the activity of the studied SOD.

Glutathione-S-transferase (GST) is an enzyme that partici-pates in the detoxification process due to conjugation reactionbetween GSH and xenobiotics Adang et al. (1990). In consis-

tence with the results of Yousefipour et al. (2005), the presentstudy indicates a significant increase in GST activity followingCP administration. An increase in GST activity might be aphysiological response to increased free radical level. Since

glutathione is utilized by the body to detoxify xenobioticsand remove free radicals through its direct antioxidant effect,the toxic levels of acrolein stimulate induction of GST activity,

thereby depleting GSH and causing direct damage to thevasculature due to free radical accumulation Yousefipouret al. (2005). However, treatment with MPS normalized the

GST level through its antioxidant effect that has the abilityto scavenge free radicals.

In conclusion, the present study showed that MPS had

positive antioxidant effect against CP-induced oxidative stressin the kidney tissues, as it alleviates the alterations in urea,creatinine and uric acid levels as well as the oxidative stressmarkers in the kidney tissues (MDA, GSH, SOD and GST).

In addition, combining MPS with silymarin showed morepronouncement effect than each one individually. It is worthto be mentioned that MPS (500 and 1000 mg/kg body weight)

and silymarin treatments restore the MDA and GSH nearly tocontrol levels.

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