Journal of Ethnopharmacology 146 (2013) 411–416

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Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jep

Efficacy of Adiantum capillus veneris Linn in chemically induced

urolithiasis 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, India
b
Department of Obstetrics and Gynecology, Government Nizamia Tibbiya College, Hyderabad, India

a r t i c l e i n f o abstract

Article history: Ethnopharmacological relevance: Adiantum capillus veneris Linn has been recommended in ancient
Received 16 July 2012 literature of Unani system of medicine as an important ingredient of many formulations for the
Received in revised form treatment of urolithiasis. Its decoction has long been used for the same purpose by several Unani
17 December 2012
physicians.
Accepted 8 January 2013
Aim of study: To investigate the antiurolithiasic effect of the hydro alcoholic extract of Adiantum capillus
Available online 17 January 2013
veneris Linn in male Sprague Dawley rats.
Keywords: Material and methods: The effects of oral administration of hydro alcoholic extract of test drug were
Unani medicine studied on calcium oxalate urolithiasis. A total of 48 rats were used for the study. The animals were
Urolithiasis
divided into six groups of eight animals each. Plain control rats were treated with distilled water only,
Antimicrobial
throughout the study period, whereas in other groups nephrolithiasis was induced by providing
Antiinflammatory
Calcium oxalate crystallization drinking water containing 0.75% ethylene glycol and 1% ammonium chloride for 7 days. Thereafter,
Antioxidant urine was examined for the presence of crystals. Negative control group A rats were sacrificed after
Flavonoids 7 days, whereas negative control group B was left untreated up to the end of study. Test groups were
Ethylene glycol treated with 127.6 mg/kg and 255.2 mg/kg of test drug and standard control with Cystone (750 mg/kg)
Ammonium chloride 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 (po 0.001 and po 0.01) 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 succession
of several physicochemical events including supersaturation,
nucleation, growth, aggregation and retention within the renal
tubules (Bouanani et al., 2010). Levels of urinary supersaturation
co-relate with the type of stone formed. Any cellular dysfunction
Abbreviations: AC, Ammonium chloride; ACV, Adiantum capillus veneris Linn;
BUN, Blood urea nitrogen; CaOx, Calcium oxalate; CPCSEA, Committee for the
purpose of control and supervision of experiment on animals; EG, Ethylene glycol;
GFR, Glomerular filtration rate; HPF, High power field; LPO, Lipid peroxidation
n
Correspondence to: H-No. 16-6-427, Osmanpur, Chaderghat, Hyderabad
500024, India. Mobile: þ91 9916967088; fax: þ91 033 22493816.
E-mail addresses: aziznium@gmail.com (A. Ahmed),
drwadud87@gmail.com (A. Wadud), nasreen2000@yahoo.com (N. Jahan),
aliabilal03@gmail.com (A. Bilal), Abdullahsyed71@yahoo.in (S. Hajera).
that can affect various urinary ions and other substances can also
influence calcium oxalate supersaturation and crystallization in the
kidney. Formation of renal stone starts with the transient super-
saturation that occurs within kidneys while excretion of millions of
urinary crystals through them. However, supersaturation is only
one step in the process of stone formation. It further needs crystals
to be retained and cause ulceration within the kidneys. Renal
injury in its turn, promotes crystal retention and development of a
stone nidus on the renal papillary surface and further supports
crystal nucleation at lower supersaturation. Reactive oxygen spe-
cies (ROSS) also seem to be responsible for cellular injury, therefore
a reduction of renal oxidative stress could also be an effective
therapeutic approach (Butterweck and Khan, 2009). Till date there
is no satisfactory drug to be used for clinical therapy. A number of
vegetable drugs are being used in many parts of the world for the
treatment of urolithiasis (Bouanani et al., 2010). Adiantum capillus
veneris Linn is traditionally used in Unani system of medicine for

0378-8741/$ - see front matter & 2013 Elsevier Ireland Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jep.2013.01.011
412 A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416
the treatment of inflammatory diseases (Haider et al., 2011). It is
an important drug, widely used in patients of urolithiasis and is
included as an important ingredient in many formulations, used for
litholytic activity. Its decoction is frequently used for its lithotriptic
effect and considered to be capable of expelling stones from kidney
and bladder (Ibn Sina, 2007; Ghani, 2011). Moreover, during
chemical 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 (Takahisa
et al., 1999; Pourmorad et al., 2006) which has been documented
by Yadav et al. (2011) to be responsible for litholytic activity. Some
of the studies carried out on Adiantum capillus veneris Linn include
evaluation of antifungal (Guha et al., 2005), antibacterial (Parihar
et al., 2010), antiviral (Abbasi et al., 2009) and antioxidant activities
(Pourmorad et al., 2006). There seems to be no report on the
antiurolithiasic activity of this drug. Therefore, this study was
undertaken to investigate the anti urolithiasic effect of the hydro
alcoholic extract of Adiantum capillus veneris Linn on calcium
oxalate urolithiasis, induced by ethylene glycol and ammonium
chloride in male Sprague Dawley (SD) rats.
2. Material and methods
2.1. Animals
The study was carried out on healthy male Sprague Dawley
rats weighing 180–210 g. The animals were procured from
registered 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 dark
cycle throughout the experimental period and were provided
with standard diet and water ad libitum unless stated otherwise.
The animal care procedures and experimental protocol were in
accord with the guidelines of the Committee for the Purpose of
Control and Supervision of Experiment on Animals (CPCSEA).
Study was conducted after obtaining the ethical clearance by
the Institutional Animal Ethics Committee (IAEC) of National
Institute 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. Ethylene
glycol and ammonium chloride were obtained from Sigma Aldrich
Chemicals Pvt. Limited, Bangalore, India. Cystones manufactured
by the Himalaya Drug Company, Bangalore, was purchased from
the market of Bangalore. Kits used in this study for the determi-
nation of calcium, blood urea nitrogen (BUN), creatinine, and
phosphorus 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 from
an authentic herb supplier in the local market of Bangalore, India,
identified by Dr. Siddamallayya N.A. at National Ayurveda Dietetics
Research Institute, Department of AYUSH, Ministry of Health and
Family 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 Department
of Ilmul Advia, NIUM, Bangalore for future reference. The dried plant
was made free of dirt and ground to powder using commercial mill,
100 g of which was then extracted in solvent (50% distilled water
and 50% ethanol) for 6 h in a Soxhlet apparatus at 80 1C. The extract
was filtered by filter paper (Whatman no. 40) and evaporated on
water bath till it dried completely (Karim et al., 2011). The yield of
the hydro alcoholic extract was found to be 11% w/w. The extract
was stored in a refrigerator pending the time of biological
investigations.
2.3.1. Dosage of the drug
The human therapeutic dose of Adiantum capillus veneris Linn
as mentioned in Unani classical literature is 10 g (Ghani, 2011).
The dose for rats was calculated by dividing it by adult human
weight of 60 kg and multiplying it with the conversion factor of
7 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 by
Haider et al., (2011). The dose of the extract was determined
with reference to the yield% of crude drug and found to be
127.6 mg/kg. Another higher dose was also calculated that is just
double of the first dose (255.2 mg/kg) to evaluate the efficacy of
test drug in dose-dependent manner. Fresh aqueous suspension
(in 1 ml distilled water) was prepared daily before each
administration.
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 the
animals were weighed and divided into six groups of eight
animals each. Group I served as plain control and received regular
rat food and drinking water ad libitum. The animals of groups II–
VI were treated with ethylene glycol (EG) 0.75% (V/V) and
ammonium chloride (AC) 1% (W/V) by adding in their drinking
water for 7 days for induction of urolithiasis. On the 8th day, they
were again weighed and kept in individual metabolic cages for 3 h
to collect fresh urine samples. Animals had free access to drinking
water during the urine collection period. Urine was analyzed for
crystalluria through microscopy, for which 1 ml of the fresh urine
sample was centrifuged at 3000 rpm and then 950 ml of the
supernatant was discarded. Ten microlitres of the vortex mixed
sediment was then transferred to slide. The number of the
crystals were identified and counted using light microscope
(45  ) (Fan et al., 1999). Thereafter, the animals of group IV were
treated with Cystone with a dose of 750 mg/kg (Mitra et al., 1998)
and served as standard control. The animals of group V and group
VI were treated with the hydro alcoholic extract of test drug with
a doses of 127.6 mg/kg and 255.2 mg/ kg and served as Test
groups A and B, respectively. The treatment was continued for 21
days. The animals of group II were sacrificed on the 8th day just
after urine collection and served as negative control A while the
animals in group III were left untreated for next 21 days and
served as negative control B. On 21st day of treatment the
animals of plain control, negative control B, standard and test
groups were again weighed and kept in metabolic cages for 3 h
urine collection. Crystalluria was analyzed by the previous
method.
2.5. Serum analysis
After collection of urine, rats were sacrificed under theopentone
anesthesia (50 mg/kg IP). Blood samples were collected by cardiac
puncture and serum was separated by centrifugation at 10,000 rpm
for 10 min. Creatinine, urea nitrogen, calcium and phosphorus were
assessed by using auto-analyzer and specific kits, namely serum
creatinine estimating kit, serum urea nitrogen estimating kit, serum
calcium 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 for
histopathological 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 one
way ANOVA with post hoc Tukey’s test or Kruskal Wallis test with
post-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 crystals
per HPF was counted. Administration of ethylene glycol and
Ammonium chloride to Wistar rats resulted in hyperoxyluria.
Numbers of CaOx crystals were grossly increased in negative
control group A. However, supplementation with hydro alcoholic
extract of Adiantum capillus veneris Linn in the dose of 127.6 mg/
kg significantly (po0.001) reduced the elevated level of CaOx
crystals as compared to negative control group A, which was quite
similar to the results produced by Cystone treated animals.
Significant reduction (p o0.01) was observed in test group B also.
The results clearly indicate that the test drug in both the doses
has significant lithotriptic effect (Table 1).
3.2. Serum parameters
Administration of ethylene glycol and ammonium chloride
caused almost no change and a nonsignificant elevation of
calcium levels in the animals of negative control group A and
negative control group B respectively. However, a significant
reduction (po0.05) was observed in the animals treated with
standard and single dose of test drug when compared with the
animals of plain control group. No significant alteration in the
serum phosphorus level was observed in any group. The values
were found almost similar to that in plain control. The serum urea
and creatinine levels in calculi-induced animals (Table 2; negative
control group A) were remarkably increased, indicating marked
renal damage which was restored to normal when treated with
Adiantum 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. Remarkable
reduction 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 be
increased significantly when the 0 day and 7th day (po0.01)
observations were compared (Table 3). All the animals treated
with ethylene glycol and ammonium chloride for 7 days lost
weight which was found to be restored in untreated group
(negative control group B) after 21 days. During the period of
treatment, body weight of rats treated with Cystone increased

Table 1
Effect of Adiauntum capillus veneris Linn (ACV) on urine CaOx crystals in ethylene glycol ammonium chloride induced urolithiasis by light microscopy.

Groups Treatment and dose Number of crystals

On 7th day On 21st day

Plain control Distilled water 1 ml 1.71 70.29 1.437 0.20

Negative control A EG 0.75% and AC 1% 6.38 70.50nnna
Negative control B EG 0.75% and AC 1% 3.87 70.98 4.677 0.71
Standard group EG 0.75% and AC 1% þ Cystone 750 mg/kg 4.5 70.71 2.757 0.45nnn
Test group A EG 0.75% and AC 1% ACV 127.6 mg/kgþ 3.5 70.50 2.637 0.50nnn
Test group B EG 0.75% and AC 1% þ ACV 255.2 mg/kg 3.13 70.78 3.137 0.40nn

EG, Ethylene glycol. AC, Ammonium chloride. The values are expressed as mean 7 SEM (n¼ 8 animals/group). Analyzed by One way ANOVA with post-test, Tukey: compare
all pairs of columns.
nn
Statistically significant at po 0.01.
nnn
Statistically significant at p o 0.001 when compared to negative control group A when compared to plain control group.

Table 2
Effect of Adiantum capillus veneris Linn (ACV) on serum parameters in ethylene glycol ammonium chloride induced urolithiasis.

a b c
Groups Treatment Calcium (mg/dl) Phosphorus (mg/dl) Urea (mg/dl) Creatinine (mg/dl)

Plain control Distilled water 1 ml 10.46 7 0.39 2.53 70.22 58.83 7 1.83 0.467 0.02
Negative control A EG 0.75% and AC 1% 10.207 0.55 2.89 70.48 189.60 7 80.79 1.42 7 0.43nnn
Negative control B EG 0.75% and AC 1% 11.56 7 0.24 3.04 70.47 48.45 7 1.91 0.657 0.04
Standard group EG 0.75% and AC 1% þCystone 750 mg/kg 9.13 7 0.73n 3.29 70.21 48.75 7 9.63 0.677 0.04
Test group A EG 0.75% and AC 1% þACV 127.6 mg/kg 9.26 7 0.28n 2.06 70.22 43.35 7 2.95nn 0.747 0.09
Test group B EG 0.75% and AC 1% þACV 255.2 mg/kg 10.087 0.56 2.77 70.21 52.19 7 2.69 0.757 0.03

EG, Ethylene glycol. AC, Ammonium chloride. The values are expressed as mean 7 SEM (n ¼8 animals/group). Analyzed by Kruskal Wallis test with post test, Dunn:
compare all pairs of columns. NCA—Negative control A, NCB—Negative control B, TA—Test group A, PC—Plain control, SC—Standard group.
n
p o 0.05.
nn
p o0.01.
nnn
p o0.001.
a
NC B vs. SC po 0.05, NC B vs. TA po 0.05.
b
NCA vs. TA p o0.01.
c
PC vs. NCA po 0.001.
 414 A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416

Table 3
Effect 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 194.137 4.96 244.257 6.64nn 258.63 716.65
Negative control A EG 0.75% and AC 1% 191.637 2.14 162.757 11.51
Negative control B EG 0.75% and AC 1% 195.757 3.71 164.08 7 12.56nn 229.5 7 4.63nnn
Standard group Cystone 750 mg/kg 197.5 7 2.67 169.257 6.52 250.13 75.98nnn
Test group A ACV 127.6 mg/kg 2027 3.10 210 7 8.86 252.57 75.99a
Test group B ACV 255.2 mg/kg 201.88 7 2.63 207 7 10.69 266.25 79.01a

EG, Ethylene glycol. AC, Ammonium chloride, g—gram. The values are expressed as mean 7SEM (n¼ 8 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. np o 0.05.
nn
p o0.01.
nnn
po 0.001.
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) 411–416
significantly (p o0.001). However, the treated groups also
showed much improvement (Table 3).
3.4. Histopathology
In plain control (Fig. 1a) section study of the kidney by light
microscopy showed normal structure of kidney. It was found that
the administration of ethylene glycol and ammonium chloride
caused severe damage to the medulla, glomeruli, tubules, and
interstitial spaces. Blood vessel proliferation, inflammatory cells
and moderate edema were seen in negative control group A
(Fig. 1b) while in negative control B (Fig. 1c) mild degree of
damage was observed in glomeruli, tubules, and interstitial
spaces. The damage was found to be almost recovered in standard
(Fig. 1d) and test groups (Fig. 1e and f) except mild blood vessel
proliferation, edema and glomerular damage. The damage to the
capsule, tubules, and interstitial spaces as completely recovered
and looked almost similar to plain control.
4. Discussion
Ethylene glycol alone or in combination with other drugs such
as ammonium chloride is often used to study the pathogenesis of
kidney crystal deposition. Since EG is a metabolic precursor of
oxalate, administration of EG to rats results in hyperoxalauria,
CaOx crystalluria and occasional deposition of CaOx crystals in
the kidney (Fan et al., 1999). In the present study, rats were
treated with 0.75% EG and 1% AC for 7 days. The findings of the
test demonstrated that Adiantum capillus veneris Linn produced
significant lithotriptic activity which is evident by the reduction
in size of stones and crystalluria. In group of animals that received
chemicals for 7 days (after that they were sacrificed), calcium
oxalate stones in their urine were formed in good quantity and
size of the stone was also large, which almost dissolved after the
administration of the test drug. It indicates that the test drug
broke the calcium particle into subtle constituents which were
not visible, or prevented the aggregation and thereby the forma-
tion of stones. Since EG is reported to be nephrotoxic, renal
function was assessed out at the end of the study by estimating
the serum creatinine, serum urea, serum calcium, and serum
phosphorus levels (Betanabhatala et al., 2009). Increased urinary
calcium is a factor favoring the nucleation and precipitation of
calcium oxalate or apatite (calcium phosphate) from urine and
subsequent crystal growth (Soundarajan et al., 2006; Bahuguna
et al., 2009). Standard and test group A showed significant
(po0.05) reduction in serum calcium when compared to negative
control B. But the test drug at higher dose produced almost equal
result as in plain control. Similar findings were observed in case of
serum phosphorous. Increased urinary phosphorus excretion
along with the oxalate stress seems to provide an environment
appropriate for stone formation by forming calcium phosphate
crystals, which epitaxially induces calcium oxalate deposition
(Soundarajan et al., 2006). In urolithiasis, the glomerular filtration
rate (GFR) decreases due to stones in the urinary system obstruct-
ing urine flow. This leads to the accumulation of waste products
in the blood, particularly nitrogenous substances such as urea,
creatinine and uric acid. In addition, increased lipid peroxidation
and decreased levels of antioxidant potentials have been reported
in 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 with
polyunsaturated fatty acids in cell membranes (Touhami et al.,
2007; Bahuguna et al., 2009).
Remarkable reduction was observed in the serum creatinine
level of standard and both test groups clearly indicating the
415
efficacy and curative effect of test drug. AC in combination with
EG, caused CaOx depositions in the kidneys of all rats within 4–7
days. However, when the dose of ammonium chloride (AC) was
Z0.75%, the rats became sick, drank less water, and lost body
weight after 4–5 days of treatment (Fan et al., 1999). In the
present study, the body weight of animals on the 7th day was
found to be decreased in all the groups. But, after treatment of
test drug a remarkable increase (p o0.001) in the body weight
was observed. The progressive increase in the body weight also
supports the efficacy of the test drug.
It must be emphasized that a deficit in the crystallization
inhibitory effect of urine and the presence of promoters are
considered to be the most important risk factors in the process
of urinary stone disease (Selvam et al., 2001). When these
conditions favor stone formation, the antiadherent layer of
glycosaminoglycans acts as a protective barrier against urinary
stone disease. If this layer is damaged, i.e. as a consequence of
bacterial attack, a stone nucleus might develop leading to a full
stone in the urinary tract. At this point, a drug that shows
antimicrobial properties can be considered anti lithogenic by
protecting the antiadherent glycosaminoglycans layer covering
the epithelium of the collecting system (Ramezani et al., 2009).
Since the test drug has been reported to act as antimicrobial
agent against a number of bacterial strains as well as fungi,
therefore its antimicrobial activity may be considered as one of
the possible mechanisms that the test drug evolves as part of its
antilithogenic activity (Singh and Usha, 1991). Guha et al.,
2005; Parihar et al., 2010 Adiantum capillus veneris Linn has
also been reported to be an antiinflammatory agent (Haider
et al., 2011) by few workers; hence, its antimicrobial activity
along with antiinflammatory activity can be further evaluated
for 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 in
negative control group A, while in negative control B very mild
destruction were found. The findings are in favor of autoheal-
ing. Whereas, administration of Cystone and the test drug
brought the tissues to almost normal state except mild blood
vessel proliferation, edema and glomerular damage, other
features simulated plain control.
In urolithiasis, oxalate has been reported to induce lipid
peroxidation (LPO). Both in vivo and in vitro studies have revealed
that the mechanism of induction of LPO by oxalate may be
involved through the inhibition of catalase activity. It has been
further reported that the conditions which enhance peroxidation
and depletion of thiol content increase the oxalate binding
activity, which in turn, promotes nucleation and aggregation
property of stone matrix protein fractions. This behavior is also
associated with peroxidized mitochondria and nuclei suggesting
that the peroxidation can be a causative factor for the initiation of
stone formation (Ramezani et al., 2009). The test drug has
previously been reported for its antioxidant activity (Pourmorad
et al., 2006) suggesting that it at least partially evolves the
antioxidant mechanism to induce the antilithogenic effect.
A significant decrease in the size of urinary stone was observed
in animals treated with the plant extract. As reported by some
workers Adiantum capillus veneris Linn contains flavonoids; the
antiurolithiasic activity may also be because of this constituent
(Takahisa et al., 1999; Pourmorad et al., 2006). However, these
can be isolated and further investigated for the confirmation of
lithotriptic activity. The different mechanisms proposed by dif-
ferent workers such as antiinflammatory, antioxidant, antioxalu-
ric and anticalciuric activities may potentially contribute in the
process of lithotriptic effect of the test drug by inhibiting the
lithogenesis.
416 A. Ahmed et al. / Journal of Ethnopharmacology 146 (2013) 411–416
5. Conclusion
In the light of findings and discussion, it can be concluded that
the hydro alcoholic extract of Adiantum capillus veneris Linn
reduced super saturation and the size of the particles. This
property of the test drug is therefore advantageous in preventing
urinary stone formation by inducing excretion of small particles
from the kidney and reducing the chance of retention in urinary
tract. The antiinflammatory, antimicrobial and antioxidant activ-
ities may partially contribute in the process of lithotriptic effect
by inhibiting lithogenesis. Our study is in consonance with the
studies which reported the presence of flavonoids to be respon-
sible for the lithotriptic activity of herbal drugs. Further studies
are needed to clarify the exact mechanism underlying the
litholytic effect.
Acknowledgment
Authors are extremely thankful to Professor M.A. Jafri, Director,
NIUM for providing best possible facilities for smooth proceeding
of the research and Dr. G. Sofi Reader, Dept. of Ilmul Advia, NIUM
for his contribution in statistical analysis.
References
Abbasi, A.M., Khan, M.A., Ahmad, M., Zafar, M., Khan, H., Muhammad, N., Sultana,
S., 2009. Medicinal plants used for the treatment of jaundice and hepatitis
based on socioeconomic documentation. African Journal of Biotechnology 8,
1643–1650.
Bahuguna, Y.M., Rawat, M.S.M., Juyal, V., Gananarajan, G., 2009. Antilithiatic effect
of grains of Eleusine coracana. Saudi Pharmaceutical Journal 17, 182–188.
Betanabhatala, K.S., Christina, A.J.M., Sundar, B.S., Selvakumar, S., Saravanan, K.S.,
2009. Antilithiatic activity of Hibiscus Sabdariffa Linn on ethylene glycol
induced lithiasis in rats. Natural Product Radiance 8, 43–47.
Bouanani, S., Henchiri, C., Migianu Griffoni, E., Aouf, N., Lecouve, M., 2010.
Pharmacological and toxicological effects of Paronychia argentea in experi-
mental calcium oxalate nephrolithiasis in rats. Journal of Ethnopharmacology
129, 38–45.
Butterweck, V., Khan, S.R., 2009. Herbal medicines in the management of
urolithasis: alternative or complementary. Planta Medica 75, 1095–1103.
Fan, J., Micheal, A., Paramjit, G., Chandhoke, S., 1999. Impact of Ammonium
chloride administration on rat ethylene glycol urolithiasis models. Scanning
Microscopy 13, 299–306.
Freirich, E.J., Gehan, E.A., Rall, D.P., Schmidt, Skipper, H.E., 1968. Quantitative
comparison of toxicity of anti cancer agents in mouse, rat, dog, monkey and
man. Cancer Chemotherapy Report 50, 219–244.
Ghani, M.N., 2011. Khazainul Advia (reprint). Idara Kitab al Shifa, New Delhi,
pp. 1360–1361.
Guha, P., Mukhopadhyay, R., Gupta, K., 2005. Antifungal activity of the extracts and
extracted phenols from gametophytes and sporophytes of two species of
Adiuntum. Taiwania 50, 272–283.
Haider, S., Nazreen, S., Alam, M.M., Gupta, A., Hamid, H., Alam, M.S., 2011. Anti
inflammatory and anti nociceptive activities of hdroalcoholic extract and its
various fractions from Adiantum capillus veneris Linn. Journal of Ethnophar-
macology 138, 741–747.
Ibn, Sina, 2007. Al Qanoon Fit Tib (Urdu Translation by Ghulam Hussain Kantoori).
Idara Kitabul Shifa, New Delhi, pp. 212–229.
Ibraheim, Z.Z., Ahmed, A.S., Gouda, Y.G., 2011. Phytochemical and biological
studies of Adiantum capillus veneris L. Saudi Pharmaceutical Journal 7, 1–10.
Karim, M.S., Ashraf, N., Kalam, A., Jahan, N., Jafri, M.A., Ahmad, G., 2011. Effect of
Biskhapra Trianthema portulacastrum Linn leaves in adriamycin induced
nephritic syndrome. International Journal of Green Pharmacy 5, 329–335.
Mitra, S.K., Gopumadhavan, S., Venkatarangannna, M.V., Sundaram, R., 1998. Effect
of cystone, an herbal formulation on glycolic acid induced urolithiasis in rats.
Phytotherapy Research 12, 372–374.
Parihar, P., Parihar, L., Achaleshwar, B., 2010. In vitro antibacterial activity of
fronds (leaves) of some important pteridophytes. Journal of Microbiology and
Antimicrobials 2, 19–22.
Pourmorad, F., Hosseinimehr, S.J., Shahabimajd, N., 2006. Antioxidant activity,
phenol and flavonoid contents of some selected Iranian medicinal plants.
African Journal of Biotechnology 5, 1142–1145.
Ramezani, M., Nasri, S., Yassa, N., 2009. Anti nociceptive and anti inflammatory
effects of isolated fractions from Apium graveolens Linn seeds in mice.
Pharmaceutical Biology 47, 740–743.
Selvam, R., Kalaiselvi, P., Govindraj, A., Murugan, B.V., Satish, K.A.S., 2001. Effect of
A. lanata leaf and Vediuppu chunnam on the urinary risk factors of calcium
oxalate urolithiasis during experimental hyperoxaluria. Pharmacological
Research 43, 89.
Singh, R.G., Usha, K., 1991. Evaluation of antilithiatic properties of Crataeva nurala.
Journal of Research and Education in Indian Medicine 10, 35–39.
Soundarajan, P., Mahesh, R., Ramesh, T., Begum, H., 2006. Effect of Aerva lanata on
calcium oxalate urolithiasis in rats. Indian Journal of Experimental Biology 44,
981–986.
Takahisa, N., Yoko, A., Kazuo, M., Yuh, I., Hiroyuki, A., Kenji, S., 1999. Fern
constituents: six new triterpenoid alcohols from Adiantum capillus veneris.
Chemical and Pharmaceutical Bulletin 47, 543–547.
Touhami, M., Laroubi, A., Elhabazi, K., Loubna, F., Zara, I., Eljahiri, Y., et al., 2007.
Lemon juice has protective activity in a rat urolithiasis model. BioMed Central
Urology 7, 1–10.
Yadav, R.D., Jain, S.K., Alok, S., Alok, M., Bharti, J.P., Jaiswal, M., 2011. Herbal plants
used in the treatment of urolithiasis, a review. International Journal of
Pharmaceutical Science Research 2, 1412–4120.