Journal of Ethnopharmacology 146 (2013) 411–416

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Efficacy of Adiantum capillus veneris Linn in chemically inducedurolithiasis in rats

Ajij Ahmed a,n, Abdul Wadud a, Nasreen Jahan a, Alia Bilal a, Syeda Hajera b

a Department of Ilmul Avia (Pharmacology), National Institute of Unani Medicine, Kottigepalaya, Magadi Main Road, Bangalore 560091, Indiab Department of Obstetrics and Gynecology, Government Nizamia Tibbiya College, Hyderabad, India

a r t i c l e i n f o

Article history:

Received 16 July 2012

Received in revised form

17 December 2012

Accepted 8 January 2013Available online 17 January 2013

Keywords:

Unani medicine

Urolithiasis

Antimicrobial

Antiinflammatory

Calcium oxalate crystallization

Antioxidant

Flavonoids

Ethylene glycol

Ammonium chloride

41/$ - see front matter & 2013 Elsevier Irelan

x.doi.org/10.1016/j.jep.2013.01.011

viations: AC, Ammonium chloride; ACV, Adi

ood urea nitrogen; CaOx, Calcium oxalate; CP

of control and supervision of experiment on

omerular filtration rate; HPF, High power fiel

espondence to: H-No. 16-6-427, Osmanp

, India. Mobile: þ91 9916967088; fax: þ91 0

ail addresses: aziznium@gmail.com (A. Ahme

d87@gmail.com (A. Wadud), nasreen2000@y

03@gmail.com (A. Bilal), Abdullahsyed71@ya

a b s t r a c t

Ethnopharmacological relevance: Adiantum capillus veneris Linn has been recommended in ancient

literature of Unani system of medicine as an important ingredient of many formulations for the

treatment of urolithiasis. Its decoction has long been used for the same purpose by several Unani

physicians.

Aim of study: To investigate the antiurolithiasic effect of the hydro alcoholic extract of Adiantum capillus

veneris Linn in male Sprague Dawley rats.

Material and methods: The effects of oral administration of hydro alcoholic extract of test drug were

studied on calcium oxalate urolithiasis. A total of 48 rats were used for the study. The animals were

divided into six groups of eight animals each. Plain control rats were treated with distilled water only,

throughout the study period, whereas in other groups nephrolithiasis was induced by providing

drinking water containing 0.75% ethylene glycol and 1% ammonium chloride for 7 days. Thereafter,

urine was examined for the presence of crystals. Negative control group A rats were sacrificed after

7 days, whereas negative control group B was left untreated up to the end of study. Test groups were

treated with 127.6 mg/kg and 255.2 mg/kg of test drug and standard control with Cystone (750 mg/kg)

for 21 days. At the end of experiment, number of crystals in urine and levels of calcium, phosphorus,

urea and creatinine in serum were observed. Histopathological study of the kidney was done by light

microscopy.

Results: Urine microscopy showed significant reduction (po0.001 and po0.01) in the number of

crystals in test groups A and B respectively. Serum levels of calcium, phosphorous, and blood urea were

found to be decreased significantly in all the groups. In both the test groups, serum creatinine level was

found to be similar as in plain control. The animals treated with test drug showed much improvement

in body weight. Histopathology of kidney showed almost normal kidney architecture in treated groups.

Conclusion: The above findings indicate the antiurolithic activity of Adiantum capillus veneris Linn, and

thus, validate the claims of Unani physicians for its medicinal use in urolithiasis.

& 2013 Elsevier Ireland Ltd. All rights reserved.

1. Introduction

Urolithiasis is a complex process that results from a successionof several physicochemical events including supersaturation,nucleation, growth, aggregation and retention within the renaltubules (Bouanani et al., 2010). Levels of urinary supersaturationco-relate with the type of stone formed. Any cellular dysfunction

d Ltd. All rights reserved.

antum capillus veneris Linn;

CSEA, Committee for the

animals; EG, Ethylene glycol;

d; LPO, Lipid peroxidation

ur, Chaderghat, Hyderabad

33 22493816.

d),

ahoo.com (N. Jahan),

hoo.in (S. Hajera).

that can affect various urinary ions and other substances can alsoinfluence calcium oxalate supersaturation and crystallization in thekidney. Formation of renal stone starts with the transient super-saturation that occurs within kidneys while excretion of millions ofurinary crystals through them. However, supersaturation is onlyone step in the process of stone formation. It further needs crystalsto be retained and cause ulceration within the kidneys. Renalinjury in its turn, promotes crystal retention and development of astone nidus on the renal papillary surface and further supportscrystal nucleation at lower supersaturation. Reactive oxygen spe-cies (ROSS) also seem to be responsible for cellular injury, thereforea reduction of renal oxidative stress could also be an effectivetherapeutic approach (Butterweck and Khan, 2009). Till date thereis no satisfactory drug to be used for clinical therapy. A number ofvegetable drugs are being used in many parts of the world for thetreatment of urolithiasis (Bouanani et al., 2010). Adiantum capillus

veneris Linn is traditionally used in Unani system of medicine for

A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416412

the treatment of inflammatory diseases (Haider et al., 2011). It isan important drug, widely used in patients of urolithiasis and isincluded as an important ingredient in many formulations, used forlitholytic activity. Its decoction is frequently used for its lithotripticeffect and considered to be capable of expelling stones from kidneyand bladder (Ibn Sina, 2007; Ghani, 2011). Moreover, duringchemical analysis, it is observed that beside tritepenes, phenylpro-panoids and carotenoids (Haider et al., 2011; Ibraheim et al., 2011)Adiantum capillus veneris Linn, also possesses flavonoid (Takahisaet al., 1999; Pourmorad et al., 2006) which has been documentedby Yadav et al. (2011) to be responsible for litholytic activity. Someof the studies carried out on Adiantum capillus veneris Linn includeevaluation of antifungal (Guha et al., 2005), antibacterial (Pariharet al., 2010), antiviral (Abbasi et al., 2009) and antioxidant activities(Pourmorad et al., 2006). There seems to be no report on theantiurolithiasic activity of this drug. Therefore, this study wasundertaken to investigate the anti urolithiasic effect of the hydroalcoholic extract of Adiantum capillus veneris Linn on calciumoxalate urolithiasis, induced by ethylene glycol and ammoniumchloride in male Sprague Dawley (SD) rats.

2. Material and methods

2.1. Animals

The study was carried out on healthy male Sprague Dawleyrats weighing 180–210 g. The animals were procured fromregistered breeder and allowed to get acclimatized for 1 week.They were housed in clean polypropylene cages at room tem-perature (2572 1C), humidity 45–55% with 12 h light–12 h darkcycle throughout the experimental period and were providedwith standard diet and water ad libitum unless stated otherwise.The animal care procedures and experimental protocol were inaccord with the guidelines of the Committee for the Purpose ofControl and Supervision of Experiment on Animals (CPCSEA).Study was conducted after obtaining the ethical clearance bythe Institutional Animal Ethics Committee (IAEC) of NationalInstitute of Unani Medicine (NIUM), Bangalore, India. (Reg. no.IAEC/VII/01/IA).

2.2. Chemicals and reagents

All the chemicals used were of analytical grade. Ethyleneglycol and ammonium chloride were obtained from Sigma AldrichChemicals Pvt. Limited, Bangalore, India. Cystones manufacturedby the Himalaya Drug Company, Bangalore, was purchased fromthe market of Bangalore. Kits used in this study for the determi-nation of calcium, blood urea nitrogen (BUN), creatinine, andphosphorus were purchased from Lab Care Diagnostics (India)Pvt. Ltd., Bangalore.

2.3. Plant materials and preparation of extract

The fronds of Adiantum capillus veneris Linn were purchased froman authentic herb supplier in the local market of Bangalore, India,identified by Dr. Siddamallayya N.A. at National Ayurveda DieteticsResearch Institute, Department of AYUSH, Ministry of Health andFamily Welfare, Government of India, Ashoka Pillar, Jayanagar,Bangalore. A voucher specimen (Ref. no. Drug Authentication/SMPU/NADRI/BNG/2010-11/45a.) was deposited in the Departmentof Ilmul Advia, NIUM, Bangalore for future reference. The dried plantwas made free of dirt and ground to powder using commercial mill,100 g of which was then extracted in solvent (50% distilled waterand 50% ethanol) for 6 h in a Soxhlet apparatus at 80 1C. The extractwas filtered by filter paper (Whatman no. 40) and evaporated on

water bath till it dried completely (Karim et al., 2011). The yield ofthe hydro alcoholic extract was found to be 11% w/w. The extractwas stored in a refrigerator pending the time of biologicalinvestigations.

2.3.1. Dosage of the drug

The human therapeutic dose of Adiantum capillus veneris Linnas mentioned in Unani classical literature is 10 g (Ghani, 2011).The dose for rats was calculated by dividing it by adult humanweight of 60 kg and multiplying it with the conversion factor of7 to accommodate the surface area of animal (Freirich et al., 1968)and found to be 1.16 gm/kg which was lesser than the safe dose,3 gm/kg, as indicated by acute toxicity study carried out byHaider et al., (2011). The dose of the extract was determinedwith reference to the yield% of crude drug and found to be127.6 mg/kg. Another higher dose was also calculated that is justdouble of the first dose (255.2 mg/kg) to evaluate the efficacy oftest drug in dose-dependent manner. Fresh aqueous suspension(in 1 ml distilled water) was prepared daily before eachadministration.

2.4. Ethylene glycol and ammonium chloride induced urolithiasis

This test was carried out by the method of Fan et al. (1999)with some modification in the treatment schedule. All theanimals were weighed and divided into six groups of eightanimals each. Group I served as plain control and received regularrat food and drinking water ad libitum. The animals of groups II–VI were treated with ethylene glycol (EG) 0.75% (V/V) andammonium chloride (AC) 1% (W/V) by adding in their drinkingwater for 7 days for induction of urolithiasis. On the 8th day, theywere again weighed and kept in individual metabolic cages for 3 hto collect fresh urine samples. Animals had free access to drinkingwater during the urine collection period. Urine was analyzed forcrystalluria through microscopy, for which 1 ml of the fresh urinesample was centrifuged at 3000 rpm and then 950 ml of thesupernatant was discarded. Ten microlitres of the vortex mixedsediment was then transferred to slide. The number of thecrystals were identified and counted using light microscope(45� ) (Fan et al., 1999). Thereafter, the animals of group IV weretreated with Cystone with a dose of 750 mg/kg (Mitra et al., 1998)and served as standard control. The animals of group V and groupVI were treated with the hydro alcoholic extract of test drug witha doses of 127.6 mg/kg and 255.2 mg/ kg and served as Testgroups A and B, respectively. The treatment was continued for 21days. The animals of group II were sacrificed on the 8th day justafter urine collection and served as negative control A while theanimals in group III were left untreated for next 21 days andserved as negative control B. On 21st day of treatment theanimals of plain control, negative control B, standard and testgroups were again weighed and kept in metabolic cages for 3 hurine collection. Crystalluria was analyzed by the previousmethod.

2.5. Serum analysis

After collection of urine, rats were sacrificed under theopentoneanesthesia (50 mg/kg IP). Blood samples were collected by cardiacpuncture and serum was separated by centrifugation at 10,000 rpmfor 10 min. Creatinine, urea nitrogen, calcium and phosphorus wereassessed by using auto-analyzer and specific kits, namely serumcreatinine estimating kit, serum urea nitrogen estimating kit, serumcalcium estimating kit and serum phosphorus estimating kit,manufactured by Lab Care Diagnostics (India) Pvt. Ltd., Bangalore.One kidney from one animal of each group was dissected out,

A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416 413

washed with tap water and preserved in 10% formalin solution forhistopathological studies (Touhami et al., 2007).

2.6. Statistical analysis

Results were expressed as mean7standard error of the mean(SEM). The results among different groups were analyzed by oneway ANOVA with post hoc Tukey’s test or Kruskal Wallis test withpost-hoc Dunn’s test. Statistical difference was considered sig-nificant at po0.05.

3. Results

3.1. Urine analysis

Fresh urine was collected for 3 h and the number of crystalsper HPF was counted. Administration of ethylene glycol andAmmonium chloride to Wistar rats resulted in hyperoxyluria.Numbers of CaOx crystals were grossly increased in negativecontrol group A. However, supplementation with hydro alcoholicextract of Adiantum capillus veneris Linn in the dose of 127.6 mg/kg significantly (po0.001) reduced the elevated level of CaOxcrystals as compared to negative control group A, which was quitesimilar to the results produced by Cystone treated animals.Significant reduction (po0.01) was observed in test group B also.The results clearly indicate that the test drug in both the doseshas significant lithotriptic effect (Table 1).

Table 1Effect of Adiauntum capillus veneris Linn (ACV) on urine CaOx crystals in ethylene glyc

Groups Treatment and dose

Plain control Distilled water 1 ml

Negative control A EG 0.75% and AC 1%

Negative control B EG 0.75% and AC 1%

Standard group EG 0.75% and AC 1% þCystone 750 mg/k

Test group A EG 0.75% and AC 1% ACV 127.6 mg/kgþ

Test group B EG 0.75% and AC 1% þACV 255.2 mg/kg

EG, Ethylene glycol. AC, Ammonium chloride. The values are expressed as mean7SEM (

all pairs of columns.nn Statistically significant at po0.01.nnn Statistically significant at po0.001 when compared to negative control group A

Table 2Effect of Adiantum capillus veneris Linn (ACV) on serum parameters in ethylene glycol

Groups Treatment aCalcium (m

Plain control Distilled water 1 ml 10.4670.3

Negative control A EG 0.75% and AC 1% 10.2070.5

Negative control B EG 0.75% and AC 1% 11.5670.2

Standard group EG 0.75% and AC 1%þCystone 750 mg/kg 9.1370.7

Test group A EG 0.75% and AC 1%þACV 127.6 mg/kg 9.2670.2

Test group B EG 0.75% and AC 1%þACV 255.2 mg/kg 10.0870.5

EG, Ethylene glycol. AC, Ammonium chloride. The values are expressed as mean7SE

compare all pairs of columns. NCA—Negative control A, NCB—Negative control B, TA—

n po0.05.nn po0.01.nnn po0.001.a NC B vs. SC po0.05, NC B vs. TA po0.05.b NCA vs. TA po0.01.c PC vs. NCA po0.001.

3.2. Serum parameters

Administration of ethylene glycol and ammonium chloridecaused almost no change and a nonsignificant elevation ofcalcium levels in the animals of negative control group A andnegative control group B respectively. However, a significantreduction (po0.05) was observed in the animals treated withstandard and single dose of test drug when compared with theanimals of plain control group. No significant alteration in theserum phosphorus level was observed in any group. The valueswere found almost similar to that in plain control. The serum ureaand creatinine levels in calculi-induced animals (Table 2; negativecontrol group A) were remarkably increased, indicating markedrenal damage which was restored to normal when treated withAdiantum capillus veneris Linn extract at the dose of 127.6 mg/kg.However, Cystone and the test drug extract in higher dose(255.2 mg/kg) produced a nonsignificant reduction. Remarkablereduction in untreated group (Table 2; negative control group B)may be due to autohealing process.

3.3. Body weight

In plain control group the body weight was found to beincreased significantly when the 0 day and 7th day (po0.01)observations were compared (Table 3). All the animals treatedwith ethylene glycol and ammonium chloride for 7 days lostweight which was found to be restored in untreated group(negative control group B) after 21 days. During the period oftreatment, body weight of rats treated with Cystone increased

ol ammonium chloride induced urolithiasis by light microscopy.

Number of crystals

On 7th day On 21st day

1.7170.29 1.4370.20

6.3870.50nnna

3.8770.98 4.6770.71

g 4.570.71 2.7570.45nnn

3.570.50 2.6370.50nnn

3.1370.78 3.1370.40nn

n¼8 animals/group). Analyzed by One way ANOVA with post-test, Tukey: compare

when compared to plain control group.

ammonium chloride induced urolithiasis.

g/dl) Phosphorus (mg/dl) bUrea (mg/dl) cCreatinine (mg/dl)

9 2.5370.22 58.8371.83 0.4670.02

5 2.8970.48 189.60780.79 1.4270.43nnn

4 3.0470.47 48.4571.91 0.6570.04

3n 3.2970.21 48.7579.63 0.6770.04

8n 2.0670.22 43.3572.95nn 0.7470.09

6 2.7770.21 52.1972.69 0.7570.03

M (n¼8 animals/group). Analyzed by Kruskal Wallis test with post test, Dunn:

Test group A, PC—Plain control, SC—Standard group.

Table 3Effect of Adiantum capillus veneris Linn (ACV) on rat body weight in ethylene glycol ammonium chloride induced urolithiasis.

Groups Treatment Weight in grams

0 day 7th day 21st day

Plain control Distilled water 1 ml 194.1374.96 244.2576.64nn 258.63716.65

Negative control A EG 0.75% and AC 1% 191.6372.14 162.75711.51

Negative control B EG 0.75% and AC 1% 195.7573.71 164.08712.56nn 229.57 4.63nnn

Standard group Cystone 750 mg/kg 197.572.67 169.2576.52 250.1375.98nnn

Test group A ACV 127.6 mg/kg 20273.10 21078.86 252.5775.99a

Test group B ACV 255.2 mg/kg 201.8872.63 207710.69 266.2579.01a

EG, Ethylene glycol. AC, Ammonium chloride, g—gram. The values are expressed as mean7SEM (n¼8 animals/group). Analyzed by ANOVA with post test, Tukey: compare

all pairs of columns. All the groups were compared with 0 day vs. 7th day, 7th day vs. 21st day. npo0.05.nn po0.01.nnn po0.001.a 0 day vs. 21st day, 7th day vs. 21st day.

Fig. 1. Microscopic images (10� ) of kidney sections after Haematoxylin and Eosin staining from (a) plain control rat showing normal epithelial lining and tubules.

Sections from (b) negative control A and (c) negative control B show severe and mild degree of damage to the medulla, glomeruli, tubules and interstitial spaces. Mild

blood vessel proliferation, edema and glomerular damage can be seen in sections from (d) standard group (e) and (f) test groups treated with Adiantum capillus veneris Linn

(ACV) extract, 127.6 and 255.2 mg/kg respectively.

A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416414

A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416 415

significantly (po0.001). However, the treated groups alsoshowed much improvement (Table 3).

3.4. Histopathology

In plain control (Fig. 1a) section study of the kidney by lightmicroscopy showed normal structure of kidney. It was found thatthe administration of ethylene glycol and ammonium chloridecaused severe damage to the medulla, glomeruli, tubules, andinterstitial spaces. Blood vessel proliferation, inflammatory cellsand moderate edema were seen in negative control group A(Fig. 1b) while in negative control B (Fig. 1c) mild degree ofdamage was observed in glomeruli, tubules, and interstitialspaces. The damage was found to be almost recovered in standard(Fig. 1d) and test groups (Fig. 1e and f) except mild blood vesselproliferation, edema and glomerular damage. The damage to thecapsule, tubules, and interstitial spaces as completely recoveredand looked almost similar to plain control.

4. Discussion

Ethylene glycol alone or in combination with other drugs suchas ammonium chloride is often used to study the pathogenesis ofkidney crystal deposition. Since EG is a metabolic precursor ofoxalate, administration of EG to rats results in hyperoxalauria,CaOx crystalluria and occasional deposition of CaOx crystals inthe kidney (Fan et al., 1999). In the present study, rats weretreated with 0.75% EG and 1% AC for 7 days. The findings of thetest demonstrated that Adiantum capillus veneris Linn producedsignificant lithotriptic activity which is evident by the reductionin size of stones and crystalluria. In group of animals that receivedchemicals for 7 days (after that they were sacrificed), calciumoxalate stones in their urine were formed in good quantity andsize of the stone was also large, which almost dissolved after theadministration of the test drug. It indicates that the test drugbroke the calcium particle into subtle constituents which werenot visible, or prevented the aggregation and thereby the forma-tion of stones. Since EG is reported to be nephrotoxic, renalfunction was assessed out at the end of the study by estimatingthe serum creatinine, serum urea, serum calcium, and serumphosphorus levels (Betanabhatala et al., 2009). Increased urinarycalcium is a factor favoring the nucleation and precipitation ofcalcium oxalate or apatite (calcium phosphate) from urine andsubsequent crystal growth (Soundarajan et al., 2006; Bahugunaet al., 2009). Standard and test group A showed significant(po0.05) reduction in serum calcium when compared to negativecontrol B. But the test drug at higher dose produced almost equalresult as in plain control. Similar findings were observed in case ofserum phosphorous. Increased urinary phosphorus excretionalong with the oxalate stress seems to provide an environmentappropriate for stone formation by forming calcium phosphatecrystals, which epitaxially induces calcium oxalate deposition(Soundarajan et al., 2006). In urolithiasis, the glomerular filtrationrate (GFR) decreases due to stones in the urinary system obstruct-ing urine flow. This leads to the accumulation of waste productsin the blood, particularly nitrogenous substances such as urea,creatinine and uric acid. In addition, increased lipid peroxidationand decreased levels of antioxidant potentials have been reportedin the kidneys of rats supplemented with calculi producing diet.In this context, oxalate has been reported to induce lipid perox-idation and to cause renal tissue damage by reacting withpolyunsaturated fatty acids in cell membranes (Touhami et al.,2007; Bahuguna et al., 2009).

Remarkable reduction was observed in the serum creatininelevel of standard and both test groups clearly indicating the

efficacy and curative effect of test drug. AC in combination withEG, caused CaOx depositions in the kidneys of all rats within 4–7days. However, when the dose of ammonium chloride (AC) wasZ0.75%, the rats became sick, drank less water, and lost bodyweight after 4–5 days of treatment (Fan et al., 1999). In thepresent study, the body weight of animals on the 7th day wasfound to be decreased in all the groups. But, after treatment oftest drug a remarkable increase (po0.001) in the body weightwas observed. The progressive increase in the body weight alsosupports the efficacy of the test drug.

It must be emphasized that a deficit in the crystallizationinhibitory effect of urine and the presence of promoters areconsidered to be the most important risk factors in the processof urinary stone disease (Selvam et al., 2001). When theseconditions favor stone formation, the antiadherent layer ofglycosaminoglycans acts as a protective barrier against urinarystone disease. If this layer is damaged, i.e. as a consequence ofbacterial attack, a stone nucleus might develop leading to a fullstone in the urinary tract. At this point, a drug that showsantimicrobial properties can be considered anti lithogenic byprotecting the antiadherent glycosaminoglycans layer coveringthe epithelium of the collecting system (Ramezani et al., 2009).Since the test drug has been reported to act as antimicrobialagent against a number of bacterial strains as well as fungi,therefore its antimicrobial activity may be considered as one ofthe possible mechanisms that the test drug evolves as part of itsantilithogenic activity (Singh and Usha, 1991). Guha et al.,2005; Parihar et al., 2010 Adiantum capillus veneris Linn hasalso been reported to be an antiinflammatory agent (Haideret al., 2011) by few workers; hence, its antimicrobial activityalong with antiinflammatory activity can be further evaluatedfor its antilithogenic activity as found in the study. In histo-pathological study of kidney, severe damage to the glomeruli,tubules and interstitial spaces, severe blood vessel prolifera-tion, inflammatory cells and moderate edema were seen innegative control group A, while in negative control B very milddestruction were found. The findings are in favor of autoheal-ing. Whereas, administration of Cystone and the test drugbrought the tissues to almost normal state except mild bloodvessel proliferation, edema and glomerular damage, otherfeatures simulated plain control.

In urolithiasis, oxalate has been reported to induce lipidperoxidation (LPO). Both in vivo and in vitro studies have revealedthat the mechanism of induction of LPO by oxalate may beinvolved through the inhibition of catalase activity. It has beenfurther reported that the conditions which enhance peroxidationand depletion of thiol content increase the oxalate bindingactivity, which in turn, promotes nucleation and aggregationproperty of stone matrix protein fractions. This behavior is alsoassociated with peroxidized mitochondria and nuclei suggestingthat the peroxidation can be a causative factor for the initiation ofstone formation (Ramezani et al., 2009). The test drug haspreviously been reported for its antioxidant activity (Pourmoradet al., 2006) suggesting that it at least partially evolves theantioxidant mechanism to induce the antilithogenic effect.

A significant decrease in the size of urinary stone was observedin animals treated with the plant extract. As reported by someworkers Adiantum capillus veneris Linn contains flavonoids; theantiurolithiasic activity may also be because of this constituent(Takahisa et al., 1999; Pourmorad et al., 2006). However, thesecan be isolated and further investigated for the confirmation oflithotriptic activity. The different mechanisms proposed by dif-ferent workers such as antiinflammatory, antioxidant, antioxalu-ric and anticalciuric activities may potentially contribute in theprocess of lithotriptic effect of the test drug by inhibiting thelithogenesis.

A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416416

5. Conclusion

In the light of findings and discussion, it can be concluded thatthe hydro alcoholic extract of Adiantum capillus veneris Linnreduced super saturation and the size of the particles. Thisproperty of the test drug is therefore advantageous in preventingurinary stone formation by inducing excretion of small particlesfrom the kidney and reducing the chance of retention in urinarytract. The antiinflammatory, antimicrobial and antioxidant activ-ities may partially contribute in the process of lithotriptic effectby inhibiting lithogenesis. Our study is in consonance with thestudies which reported the presence of flavonoids to be respon-sible for the lithotriptic activity of herbal drugs. Further studiesare needed to clarify the exact mechanism underlying thelitholytic effect.

Acknowledgment

Authors are extremely thankful to Professor M.A. Jafri, Director,NIUM for providing best possible facilities for smooth proceedingof the research and Dr. G. Sofi Reader, Dept. of Ilmul Advia, NIUMfor his contribution in statistical analysis.

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