J Young Pharm, 2020; 12(2) Suppl: s76-s81

A multifaceted peer reviewed journal in the field of Pharmacy Original Article

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Assessment of Antiurolithiatic Potentials of Crinum asiaticum

Bulbs by in vitro and in vivo Approaches
Suman Sura1*, Venkata Suresh Kumar Solleti2
1
Research Scholar, Jawaharlal Nehru Technological University Anantapur, Ananthapuramu, Andhra Pradesh, INDIA.
2
Department of Pharmacognosy and Phytochemistry, Creative Educational Society’s College of Pharmacy, Kurnool andhra Pradesh, INDIA.

ABSTRACT
Objectives: To evaluate the antiurolithiatic potential of Crinum asiaticum showed reduction in CaOx crystal aggregation and promoted nucleation
through in vitro and in vivo methods. Materials and Methods: Ethanolic after treatment with EECA. In vivo studies also showed reduction in elevated
extract of Crinum asiaticum (EECA) subjected for phytochemical screening levels of serum creatinine, BUN, uric acid and levelsof calcium, oxalate and
and HPTLC fingerprinting. In vitro antiurolithiatic activity of EECA was phosphate in urine and kidney homogenate as compared to control group.
determined by Calcium Oxalate (CaOx) crystal nucleation and aggregation The results were supported by histopathological studies. Conclusion: The
assays. Acute toxicity studies were performed as per OECD 423 guidelines. EECA have shown significant antiurolithiatic activity by reducing calculi.
For in vivo antiurolithiatic activity 36 male wistar rats divided into six groups. Key words: Antiurolithiatic activity, Crinum asiaticum, CaOx crystal
Group I served as control, groups II to VI are administered with 0.75 % v/v aggregation, CaOx crystal nucleation, HPTLC Fingerprinting.
ethylene glycol for 28 days to induce hyperoxaluria where in group II served
Correspondence
as toxic control and group III as standard. Group IV to VI served as test and
received respective doses of EECA from 15 to 28 days. After 28 days, Mr. Suman Sura,
creatinine, BUN, uric acid was estimated. Calcium, oxalate, phosphate Research Scholar, Jawaharlal Nehru Technological University Anantapur,
were estimated in urine and kidney homogenate also subjected for histo­ Ananthapuramu-515002, Andhra Pradesh, India.
pathological studies. Results: Phytochemical screening revealed the
Phone: +91 9949521100
presence of flavonoids, terpenoids, tannins and phenolic compounds.
HPTLC fingerprinting shows the presence of 7 terpenoids and 7 flavonoids Email: [email protected]
at 540nm and 366nm respectively after derivatization. In vitro studies DOI: 10.5530/jyp.2020.12s.51

INTRODUCTION
Crinum asiaticum (Common name: Asian poison bulb) locally known lithotripsy and local calculus disruption using high-power laser are
as kesaracettu, is an evergreen herb that is widely distributed throughout largely used to eliminate the calculi. nevertheless, these are costly and
India along river beds and also in forest.1 It is known as spider lily, with these procedures’ recurrence is quite common. The recurrence rate
Crinum lily and poison bulb in English, naagadamani in Ayurveda, without preventive treatment is approximately 10% at 1 year, 33% at
bakong in Malaysia and morabau in Papua New Guinea. The plant has 5 years and 50% at 10 years. The other therapies may also be involving
various ethnomedicinal properties and is used in traditional system of thiazide diuretics and alkali-citrate are used in an endeavor to prevent
medicine. The ethnomedicinal uses of the Crinum asiaticum bulbs are repetition but scientific verification for their worth is less persuasive.7
bitter, expectorant, laxative, carminative, anthelmintic, aphrodisiac,
diuretic, urinary problems, diaphoretic, nauseant,1 analgesic and anti- MATERIALS AND METHODS
inflammatory,2 anti-obesity,3 emetic,4 antitumor.5 Collection of plant material
The stone formation in the urinary system, i.e. in the kidney, ureter, The Crinum asiaticum plant was collected from the Tirumala hills, Chittoor
urinary bladder and in the urethra is known as urolithiasis. ‘Urolithiasis’ district Andhra Pradesh, India and confirmed by Dr. K. Madhava Chetty,
derived from, ouron also called urine and litho means stone. It is one a Assistant Professor, Department of Botany, Sri Venkateshwara University,
foremost disease of the urinary tract and is a basis of morbidity, formation Tirupati (Voucher Number: 2011, dated 08.08.2017). The whole plant
of stone the kind of urologic disorders it is occur in roughly 12% of was dried in shade; bulbs were separated and pulverized to get a coarse
the global population and it is occurred 70-81% in males and in females powder.
47-60%.6
Urolithiasis is a universal trouble, cautious no environmental, artistic Preparation of extract
or ethnic groups. Since before two decades incident of primary bladder The powered material was subjected for extraction by using ethanol as
stones has extensively reduced, but these are even now a day also reported solvent by soxhlation process.
in the developing world mostly in patients who are suffered with
neurogenic bladders and benign prostatic hypertrophy. Around 80% Preliminary phytochemical screening of extract
of the stones are composed of calcium oxalate (CaOx) and calcium Crinum asiaticum bulb extracts were subjected to phytochemical testing
phosphate. Urinary stones may source for the hindrance, hydronephrosis, for identification of alkaloids, glycosides, tannins, phenols, steroids,
illness and gush of blood in the urinary tract system. Surgical operation, flavonoids and terpenoids following the standard procedures.8

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 Sura and Kumar.: Assessment of Antiurolithiatic Potentials of Crinum asiaticum Bulbs

HPTLC analysis of ethanolic extract of Crinum asiaticum Evaluation of in vivo pharmacological studies
bulbs Healthy wistar albino rats weighing about 150-180 g are procured from
The ethanolic extract of Crinum asiaticum bulbs were applied in a animal house, CES college of pharmacy, Kurnool. They were housed in
polypropylene cages and maintained at 27±2°C, relative humidity
concentration of 2 μl using CAMAG Automatic TLC sampler4 applicator
65 ± 10% under 12 h light/dark cycles. The animals were given standard
on precoated aluminium sheets with silica gel 60 F254 TLC plates (Merck)
diet manufactured by Nutrivet Life Sciences, Pune, India. The study
of 0.2 mm thick, 5 cm × 20 cm, used as a stationary phase. The plates
protocol was approved by the Institutional Animal Ethics Committee
were developed in the mobile phase of n-hexane: ethyl acetate (1:1) (Ref. No.: IAEC/CESCOP/2017-10) constituted in accordance guidelines
v/v for terpenoids and ethyl acetate: formic acid: acetic acid: water of the CPCSEA (Committee for the purpose of Control and Supervision
(100:11:11:26) v/v/v/v for flavonoids.9 The plates were developed in of Experiments on Animals), India.
CAMAG – Twin trough glass chamber saturated with saturated pad for
20 min, at a distance of 70 mm. After development the plates was sprayed Acute toxicity studies
with Anisaldehyde sulphuric acid reagent and Natural product reagent The acute oral toxicity study was carried out as per the OECD-423
respectively for terpenoids and flavonoids using CAMAG derivatizer. guidelines.12 One tenth of the non-median lethal dose (LD50) was taken
The tracks were scanned using CAMAG TLC Visualizer equipped with as effective dose.13
CAMAG software© VisionCATS-Server-PH, version 2.5.18262.1 at a
wavelength of 540 nm using tungsten lamp and 366 nm using mercury Antiurolithiatic activity
lamp and the finger print profiles were recorded for terpenoids and
Ethylene glycol induced urolithiasis in wistar albino rats
flavonoids respectively.
The method described by Atmani et al.14 was used for the evaluation of
Evaluation of in vitro antiurolithiatic activity antiurolithiatic activity of ethanolic extract of Crinum asiaticum bulbs
in albino rats. Thirty-six rats were divided into six groups each group
The effect of Crinum asiaticum extracts on CaOx crystallization was consisting of six rats.
determined by the measurement of turbidity changes due to the crystal
Group-1 served as normal control receives regular diet and potable water
nucleation and aggregation. The precipitation of calcium oxalate at for 28 days
37°C and pH 6.8 has been examined by the measurement of turbidity at
Group-2 served as disease control receives ethylene glycol (0.75%) in
620 nm. A spectrophotometer UV/Vis (Lab India) was used to measure
drinking water for 28 days
the turbidity of the augmentation of calcium oxalate.
Group-3 served as standard receives ethylene glycol (0.75%) water for
28 days and Cystone (750mg/kg) from 15th day-28th day
Nucleation assay
Group-4-6 served as test groups received ethylene glycol (0.75%) water
Effect of ethanolic extract of Crinum asiaticum bulbs on calcium oxalate
for 28 days and ethanolic extract of Crinum asiaticum bulb at the doses of
(CaOx) crystal formation was determined by means of nucleation assay. (100mg/kg, 200mg/kg and 400mg/kg) respectively from 15th day-28th
Individually, calcium chloride(3 mM) and sodium oxalate(0.5 mM) day.
solutions were filtered three times through the pore size of 0.22 μm filter,
from that 950 μL of calcium chloride was took and to this added 100 μL Assessment of antiurolithiatic activity
of extract at different concentrations (50 μg - 3200 μg/mL). Then add
Serum analysis
950 μL of sodium oxalate solution for initiation of crystals. Then finally
After last dose of the drug treatment, blood was collected through retro-
the solution was magnetically moved at 800 rpm with a stirring bar. The
orbital plexus under slight anaesthetic conditions. Serum was separated
temperature 37ºC was maintained. At 620 nm the solution optical
by centrifugation (Research Centrifuge, R-22, Remi India) at 10,000×g
density has been monitored. The rate of nucleation was determined with for 10 min and analyzed for creatinine, uric acid and blood urea nitrogen
the comparison of the induction time. (The delay ahead of the exterior of (BUN). Serum parameters were estimated by semi-auto analyser (Mispa
crystals that have reached a critical size then consequently become Excel Chemistry analyser) with diagnostic kits of Excel Diagnostic Pvt.
optically demonstrable) if the control with the presence of extract no Ltd, Hyderabad.
need to addition of corm extracts.10
Urine analysis
Aggregation assay On the 28th day individual animal was kept in separate metabolic cages.
24 hr of urine sample was collected. Provide the water for rats during the
Effect of ethanolic extract of Crinum asiaticum bulbs on calcium oxalate
urine collection, urine sample was subjected for estimation of Calcium
(CaOx) crystal aggregation was determined by means of aggregation
(Calcium diagnostic Kit, Agappe Diagnostics Ltd, Kerala, India),
assay. 0.8mg/mL COM crystals were used at an ultimate concentration Oxalate15 and Phosphate.16
buffered with 0.05M Tris containing sodium chloride (0.15M) at pH 6.5.
Those all were performed at the temperature 37°C in the presence Kidney homogenate analysis and histopathology
and absence of the corm extract after the apprehend of stirring. The of At the end of the study period, the rats were euthanised by using CO2
aggregation rate was predictable as below mentioned formula, by chamber and the abdomen was cut open to remove both kidneys from
comparing the slope of the turbidity in the sample and with that turbidity each animal. Isolated kidneys were washed off extraneous tissue, rinse
in the control.11 out in ice-cold physiological saline and used for histopathology and
homogenate analysis. The left kidney was finely chopped and 20%
Turbidity of sample homogenate was prepared in Tris-Hcl buffer (0.02 mol/l, pH 7.4). Kidney
Ir = ×100
Turbidity of control homogenate was used for assaying tissue calcium, oxalate and Phosphate.
The right kidney was fixed in 10% neutral buffered formalin, treated in
Ir = Percentage aggregation inhibition rate a series of graded alcohol and xylene, fixed in paraffin wax, partitioned

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 Sura and Kumar.: Assessment of Antiurolithiatic Potentials of Crinum asiaticum Bulbs

at 5μm and stained with Hematoxylin and Eosin for examination under Table 1: HPTLC analysis of EECA.
polarized light. The histopathological examination of slides was examined Compound Terpenoids Flavonoids
under polarized light microscope (40X) and photographed by an Olympus
Rf % Area Rf % Area
Digital Camera.
1 0.070 1.76 0.160 10.09
Statistical Analysis
2 0.111 4.78 0.221 15.57
All the values are articulated as mean ± SEM. The data was statistically
3 0.287 3.59 0.302 16.78
analyzed by using one-way ANOVA followed Dunnett’s t test in GraphPad
Prism 5.03 version software. 4 0.451 21.37 0.406 45.34
5 0.592 28.24 0.519 6.00
RESULTS 6 0.790 16.57 0.826 1.37
Preliminary phytochemical screening of extract 7 0.872 23.70 0.974 4.85
Phytochemical screening of the ethanolic extract of Crinum asiaticum
bulbs revealed the presence of flavonoids, tannins, phenolics, steroids
and terpenoids.

HPTLC analysis of ethanolic extract of Crinum asiaticum

bulbs
To obtain the reproducible results with high resolution different
combinations of solvents with various ratios were tested. Satisfactory
results were obtained with n-hexane: ethyl acetate (1:1) v/v and ethyl
acetate: formic acid: acetic acid: water (100:11:11:26) v/v/v/v for
terpenoids and flavonoids respectively. The 2µl of ethanolic extract of
Crinum asiaticum bulbs showed the presence of 7 terpenoids, 7 flavonoids
with different Rf values and % of area when scanned at 540nm (Figure
1) and 366 nm (Figure 2) respectively for terpenoids and flavonoids after
derivatization. (Table 1)

In vitro antiurolithiatic activity Graph 1: Effect of different concentrations of EECA on CaOx crystal nucleation.
In the present study the different graded concentrations i.e. from 50µg/ml
to 3200 µg/ml of ethanolic extract of Crinum asiaticum bulbs were used
for in vitro evaluation of antiurolithiatic activity. The reticence of crystal
formation was directly proportional to the increase in the concentration
of EECA, with maximum activity was pragmatic at 3200 µg/ml in CaOx
crystal nucleation and aggregation assays (Graph 1,2).

Graph 2: Effect of different concentrations of EECA on CaOx crystal aggregation.

Figure 1: HPTLC chromatogram of terpenoids in 2µl of EECA.

Evaluation of in vivo pharmacological studies

Acute toxicity studies
The purified and entirely dried ethanolic extract of Crinum asiaticum
bulbs were subjected for the acute toxicity study to determine the lethal
dose using wistar albino rats in forced environment. Acute toxicity studies
were performed according to the OECD 423 guidelines. The limit test was
performed with the dose of EECA (2000 mg/kg, b. w) was administered
by oral route to a group of rats using oral feeding needle (22gauge). After
treatment to rats were monitored for 14 days and it was observed that
no changes in normal behaviour, hence it was conformed that the EECA
is virtually non-toxic in normal rats and fall under the sort of GHS
Figure 2: HPTLC chromatogram of flavonoids in 2µl of EECA. category 5, according to 1/20th dose (100 mg/kg. b.w), 1/10th of dose

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 Sura and Kumar.: Assessment of Antiurolithiatic Potentials of Crinum asiaticum Bulbs

(200 mg/kg. b.w), 1/5th dose (400 mg/kg. b.w) from maximum safe dose Table 2: Effect of ethanolic extract of Crinum asiaticum (bulbs) on serum
was considered for further evaluation pharmacological studies. levels of BUN, Creatinine and uric acid in ethylene glycol (0.75%) induced
urolithiasis in rats.
Ethylene glycol induced urolithiasis in wistar albino rats BUN Creatinine Uric acid
Serum analysis S.no Groups
(mg/dl) (mg/dl) (mg/dl)
In the present study treatment with ethylene glycol (0.75%) results in 1 Normal 28.28±1.74 0.354±0.09 1.78±0.36
a significant (###p<0.001) increase in elevated levels of (47.58±3.27,
2 Disease Control 47.58±3.27### 2.503±0.29### 4.673±0.54###
2.503±0.29,4.673±0.54) serum BUN, creatinine and uric acid respectively
when compared to normal group. These changes were restored Standard (Cystone
3 30.26±1.85*** 1.10±0.19*** 2.588±0.411*
significantly (***p<0.001; 30.26±1.85, 1.10±0.19) BUN, Creatinine 750mg/kg,BW, p.o)
and (*p<0.05; 2.588±0.411) uric acid in rats treated with the standard EECA (100 mg/kg,
4 36.06±1.63* 1.80±0.41ns 2.497±0.76*
drug (cystone 750mg/kg, p.o). However, the rats treated with EECA BW, p.o)
(100mg/kg, p.o) pointedly decreases (*p<0.05; 36.06±1.63, 2.497±0.76) EECA (200 mg/kg
in BUN, uric acid respectively and creatinine non significantly ((p>0.05; 5 34.22±3.11** 1.366±0.20* 2.530±0.49*
BW, p.o)
1.80±0.41), at the dose of EECA (200mg/kg, p.o) significantly (**p<0.01) EECA (400 mg/kg
lowers the BUN (34.22±3.11) and (*p<0.05; 1.366±0.20, 2.530±0.49) 6 31.76±2.89*** 1.20±0.33** 1.847±0.30**
BW, p.o)
creatinine and uric acid respectively, at the dose of EECA (400mg/kg, p.o)
significantly (***p<0.001) lowers the BUN (31.76±2.89) and (**p<0.01; All values are expressed as mean ±S.E.M for six rats in each group.
1.20±0.33, 1.847±0.30) creatinine and uric acid respectively when Comparisons made between ###p<0.001,##p<0.01,#p<0.05; Normal Vs Disease
compared to disease control. (Table 2). control,
***p<0.001, **p<0.01, *p<0.05; Disease control Vs Treatment: One-way ANOVA
Urine analysis followed by Dunnett’s -t test.

The administration of ethylene glycol (0.75%) to wistar albino rats triggers

Table 3: Effect of ethanolic extract of Crinum asiaticum (bulbs) on urinary
hyperoxaluria. Urinary levels of Oxalate, calcium and phosphate
levels of Calcium, oxalate and Phosphate in ethylene glycol (0.75%)
(8.16±0.39, 4.23±0.55, 7.25±0.51) were unacceptably (###p<0.001) induced urolithiasis in rats.
increased in the calculi-induced animals. The standard drug cystone
Calcium Oxalate Phosphate
(750mg/kg, p.o) treated group animals acceptably lowers the levels of S.no Groups
calcium (***p<0.001; 3.78±0.321), oxalate and phosphate (**p<0.01; (mg/dl) (mg/dl) (mg/dl)
2.26±0.20, 4.15±0.70) respectively. However, treatment with EECA 1 Normal 3.38±0.45 1.60±0.32 3.85±0.48
(100mg/kg, p.o) lowered the elevated level of calcium (**p<0.01; 2 Disease Control 8.16±0.39 ###
4.23±0.55 ###
7.25±0.51###
5.66±0.74), oxalate (*p<0.05; 2.79±0.27) and phosphate (p>0.05;
Standard (Cystone 750mg/
5.31±0.55), at the dose of EECA (200mg/kg, p.o) reduces the elevated 3 3.78±0.321*** 2.26±0.20** 4.15±0.70**
kg,BW, p.o)
levels of calcium(***p<0.001; 4.26±0.23), oxalate(*p<0.05; 2.73±0.42)
4 EECA (100 mg/kg, BW, p.o) 5.66±0.74** 2.79±0.27* 5.31±0.55ns
and phosphate(**p<0.01; 4.63±0.47), at the dose of EECA (400mg/kg,
p.o) reduces the levels of calcium and phosphate(***p<0.001; 4.05±0.59, 5 EECA (200 mg/kg BW, p.o) 4.26±0.23*** 2.73±0.42* 4.63±0.47**
4.05±0.370 and oxalate (**p<0.01; 2.35±0.18) in urine when associated 6 EECA (400 mg/kg BW, p.o) 4.05±0.59*** 2.35±0.18** 4.05±0.37***
to the disease control group (Table 3).
All values are expressed as mean ±S.E.M for six rats in each group.
Comparisons made between ###p<0.001,##p<0.01,#p<0.05; Normal Vs Disease
Kidney Homogenate analysis
control,
Supernatant from the kidney homogenate was collected and subjected ***p<0.001, **p<0.01, *p<0.05; Disease control Vs Treatment: One-way ANOVA
for evaluation of tissue calcium, oxalate and phosphate. The levels of followed by Dunnett’s -t test.
tissue calcium, oxalate and phosphate were significantly (###p<0.001)
increases (6.47±0.64###, 3.95±0.47###, 6.48±0.11###) compared to normal Table 4: Effect of ethanolic extract of Crinum asiaticum (bulbs) on Kidney
group. The animals treated with standard drug cystone (750mg/kg, p.o) homogenate levels of Calcium, oxalate and Phosphate in ethylene glycol
significantly reduced calcium (***p<0.001; 3.58±0.85), oxalate (**p<0.01; (0.75%) induced urolithiasis in rats.
2.54±0.70), phosphate (**p<0.01; 3.89±0.70) and EECA (100mg/kg, Calcium Oxalate Phosphate
p.o) significantly reduces the calcium (**p<0.01; 5.42±0.84), oxalate S.no Groups
(mg/dl) (mg/dl) (mg/dl)
(*p<0.05;3.55±0.75), phosphate (*p<0.05; 4.56±0.78), EECA (200mg/
kg, p.o) significantly lowers the calcium (***p<0.001; 4.95±0.73), oxalate 1 Normal 2.68±0.85 1.95±0.55 2.95±0.55
(*p<0.05; 3.38±0.45) and phosphate (**p<0.01; 4.05±0.50) phosphate 2 Disease Control 6.47±0.64 ###
3.95±0.47###
6.48±0.11###
respectively, EECA (400mg/kg, p.o) significantly lowers the calcium Standard (Cystone
3 3.58±0.85*** 2.54±0.70** 3.89±0.70**
(***p<0.001; 4.68±0.95), oxalate (**p<0.01; 3.68±0.74) and phosphate 750mg/kg, BW, p.o)
(***p<0.001; 3.78±0.77) correspondingly when compared to disease 4 EECA (100 mg/kg, BW, p.o) 5.42±0.84** 3.55±0.75* 4.56±0.78*
control group. (Table 4)
5 EECA (200 mg/kg BW, p.o) 4.95±0.73*** 3.38±0.45* 4.05±0.50**
Kidney Histopathology 6 EECA (400 mg/kg BW, p.o) 4.68±0.95*** 3.68±0.74** 3.78±0.77***
Histopathological examination of rat kidneys treated with ethylene All values are expressed as mean ±S.E.M for six rats in each group.
glycol (0.75% v/v) revealed the presence of calcium oxalate deposits Comparisons made between ###p<0.001,##p<0.01,#p<0.05; Normal Vs Disease
inside the renal tubules and dilation of the renal tubules along with control,
interstitial inflammation were observed. The number of calcium oxalate ***p<0.001, **p<0.01, *p<0.05; Disease control Vs Treatment: One-way ANOVA
deposits in the renal tubules and dilation of renal tubules of Groups III followed by Dunnett’s -t test.

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 Sura and Kumar.: Assessment of Antiurolithiatic Potentials of Crinum asiaticum Bulbs

out nucleation assay. The reticence of crystal formation was directly

proportional to the increase in the concentration of EECA, with maximum
activity was pragmatic at 3200 µg/ml in CaOx crystal nucleation.
This suggests the anticrystallization activity of EECA against CaOx
crystallization. One possible mechanism of anticrystallization activity of
EECA could be its proficiency to complex with free calcium and oxalate
ions, thus blocking the formation of CaOx complexes, as has also been
suggested for Sarghassum wightti.19
Aggregation is an important factor of crystal retention as large crystal
agglomerates are the ones that produce renal tubular obstruction thereby
promoting stone formation.19 EECA showed significant inhibitory effect
on CaOx crystal aggregation.
Ethylene glycol is the most commonly used inducing agent for inducing
urolithiasis in rats. The rats fed with ethylene glycol (0.75%v/v) results
significant increase in serum levels of BUN, creatinine and uric acid
and promote excessive excretion of urinary levels of calcium, oxalate
and phosphate, indicate formation of calcium oxalate stones in kidneys.
However, treatment with EECA, significantly lowers the elevated serum
levels of BUN, creatinine and uric acid and urinary levels of calcium,
oxalate and phosphate in a dose dependent manner.
Microscopic examination of rat kidney sections treated with ethylene
glycol shows the presence of calcium oxalate deposits, dilatation of renal
tubules along with interstitial inflammation. However, co treatment with
EECA reduces the number of calcium oxalate deposits, dilatation of
renal tubules and prevents the damage to renal tubules in dose dependent
manner.
The phytoconstituent found in the extract such as flavonoids, tannins,
phenolics, steroids and terpenoids may be responsible for the activity. In
earlier literature it was reported that both flavonoids20 and terpenoids21
play a significant role in antiurolithiatic activity. Attempt also made to
A-Normal group standardise the extract by performing finger printing of flavonoids and
B-Disease control (Ethylene glycol 0.75% v/v) terpenoids by HPTLC.
C-Standard (Cystone 750mg/kg, bd.wt)
D-EECA(100mg/kg, bd. wt) CONCLUSION
E-EECA(200mg/kg, bd. wt) In conclusion, the findings of the present study provide clear evidence
F-EECA(400mg/kg, bd. wt) that the EECA inhibits the development of calcium oxalate crystals in
in vitro. The oral administration of EECA to ethylene glycol induced
Figure 3: Histopathological view of the experimental groups. Sections show
urolithiasis rats’ results in reduction in the elevated serum levels of
the hematoxylin and eosin (HE) stained kidney. Sections we reviewed using
BUN, Creatinine and uric acid and urinary levels of calcium, oxalate
polarized light microscope (40X) and photographed by an Olympus Digital
Camera. and phosphate in dose dependent manner. Therefore, EECA exhibited
significant anti urolithiatic activity against ethylene glycol induced
urolithiasis in rats.
(Figure 3-C) rats was significantly less than the Group II (Figure 3-B).
Treatment with EECA with different doses (100 mg/kg, BW, p.o, ACKNOWLEDGEMENT
200 mg/kg, BW, p.o, 400 mg/kg, BW, p.o) i.e., Group IV-VI (Figure 3-D-F).
The authors would like to acknowledge monetary support from the
Significantly lowers the deposition of calcium oxalate crystal, dilation
Principal and Management, Creative Educational Society’s college of
of renal tubules and interstitial inflammation were observe compared to
Pharmacy, NH-7, Chinnatekur, Kurnool, A.P. for providing constant
Group-II (Figure 3-B).
support throughout the study.
DISCUSSION
CONFLICT OF INTEREST
CaOx urolithiasis is the most predominant type of all urinary stone
diseases. Key magnitudes concerned in its pathological bio mineralization The author claims there is no conflict of interest.
include crystal nucleation, growth and aggregation. Present study was
aimed to discourse these key events involved in CaOx stone formation as ABBREVIATIONS
a resource to investigate the efficacy of C. asiaticum bulbs as an anti­ CaOx: Calcium oxalate; COM: Calcium oxalate monohydrate; EECA:
urolithiatic agent. Nucleation is an essential in the pathogenesis of CaOx Ethanolic extract of Crinum asiaticum.
urolithiasis. Nucleation fundamentally marks a thermodynamically
driven event of phase alteration wherein dissolved substances in a REFERENCES
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Article History: Submission Date : 10-03-2020; Revised Date : 26-04-2020; Acceptance Date : 16-05-2020.
Cite this article: Sura S, Kumar SSV. Assessment of Antiurolithiatic Potentials of Crinum asiaticum Bulbs by in-vitro and in-vivo Approaches. J Young Pharm.
2020;12(2)Suppl:s76-s81.

Journal of Young Pharmacists, Vol 12, Issue 2(Suppl), Apr-Jun, 2020 S81