Antilithiatic Effect of Crataeva magna Lour. DC Leaves on Ethylene glycol induced Lithiasis

 

Sridhar N.1*, Dr. Venkataraman S.1 and Dr. Manisha Mishra S.2

1Centre for Advanced Research in Indian System of Medicine, SASTRA University, Tanjore 613 401. Tamil Nadu.

2Amity University, Lucknow 226 010, Uttar Pradesh.

 

 

ABSTRACT:

Objective: Crataeva magna is a potent medicinal plant in the Indian system of medicine.  The objective of the present work was to study the antilithic activity of plant Crataeva magna (Lour.) DC belonging to family Capparaceae.

Materials and Methods:  The present study was carried out to evaluate Antilithiatic activity in rats using 0.75% ethylene glycol in drinking water were given orally for 28 days.  The plant Crataeva magna is used in analgesic, anti protozoal, hypoglycemic, anti inflammatory, hypotensive, anti spasmodic purposes.  The aqueous extract of leaves of Crataeva magna results maximum yield value than that of petroleum ether extract, chloroform extract and alcohol extract through successive maceration process.  The aqueous extract of leaves of Crataeva magna showed maximum control of lithiasis in Wister rats.    Urinary risk factors of urolithiasis were monitored at the end of 7th , 14th , 21st , and 28th days.  Urinary volume was increased in lithiatic as well as drug-treated rats.  Increased urinary excretion of Calcium, Oxalate, Uric acid, Phosphorus and Protein in lithiatic rats was brought down significantly by the administration of Crataeva magna.  Decreased magnesium excretion in hyperoxaluric rats was normalized with respect to calcium oxalate and other crystallizing salts such as uric acid, which may induce epitaxial deposition of calcium oxalate.  Simultaneous treatment with the extract reduced Calcium and Oxalate ion concentration in Urine confirming the stone inhibitory effect.

Result: In Ethylene glycol induced lithiasis, the lithiasis was significantly reduced and the stone formation was normalized by administration of 200 mg/kg and 400 mg/kg dose orally and the property was comparable to the standard drug.

Conclusion:  This study has established the antilithiatic activity of Crataeva magna and thus, justifies the folkloric and ethnomedical uses of this plant for lithiasis.

 

KEYWORDS: Lithiasis, Ethylene glycol, Crataeva magna, rats.

 

INTRODUCTION:

Lithiasis is worldwide in distribution and a common disorder estimated to occur in approximately 12 % of the population, with a recurrence rate of 70-80% on males and 47-60% females1. Lithiasis in its different forms is frequently encountered during urological complications. Some common causes are inadequate urinary drainage, foreign bodies in the urinary tract, microbial infections, diet with excess oxalates and calcium, vitamin abnormalities, viz.  Vitamin A deficiencies, Vitamin D excess, metabolic diseases like hyperparathyroidism, cystinuria, gout and intestinal dysfunction2. Present day medical management of lithiasis mainly involves the surgical removal of stones. 

 

Techniques such as ESWL PCNL do not assure the prevention of recurrence of the stone.  They cause side effects such as hemorrhage, hypertension, tubular necrosis and subsequent fibrosis of the kidney3.

 

The present study was undertaken to evaluate the pharmacological evaluation of the effect of the plant Crataeva magna on the healing of experimentally induced lithiasis in rats.

 

MATERIALS AND METHODS:

Plant Materials:  The Leaves part of plant Crataeva magna was collected from young matured plant from Alagar Kovil, Madurai during the month of Nov-Dec and identified by the botanist of Department of Botany, American College, Madurai by comparing with the voucher specimen present in the herbarium. After authentification fresh plant materials were collected in bulk, washed under running tap water to remove adhering dust, dried under shade and pulverized in a mechanical grinder.  The coarse powder was used for further studies.

 

Preparation of Plant Extract

About 200 gm of coarse dried powder of plant of the Crataeva magna was taken in the soxhlet apparatus and extracted successively using different solvents according to their increasing order of polarity, for the present investigation.  (i.e. Pet. ether, Chloroform, Ethanol, Aqueous).  The extraction for each solvent was carried out for 18 to 24 hours.  The extract was collected by evaporating the solvents by slow heat treatment.  Total 2 kg of pulverized whole plant was subjected under solvent extraction to produce the required amount of test extract.  Calculated amount of dried aqueous extract was suspended in 0.5% w/v of sodium-CMC in normal saline solution to get the test doses (200mg/kg per ml for both extract)4.

 

Experimental animals

Wister male rats aged 16-20 weeks weighing 160-200g were obtained from the Department of  Experimental Medicine, Central Animal House, Rajah Muthiah Medical College, Annamalai University. The experimental protocol is approved by the Institutional Animal Ethics Committee, Annamalai University and animals were maintained under standard conditions for an acclimatization period of 15 days before performing the experiment.  All rats were housed individually in metabolic cages and temperature maintained at 22±2°C.

 

Ethylene glycol induced lithiasis

Animals were divided into five groups.  Each group consists of six animals.  Group I animals served as a control and were maintained using commercial pelleted feed.  Group II animals received 0.75% ethylene glycol in drinking water ad libitum for 28 days. Group III (Standard drug treated rats) were fed with  0.75% Ethylene glycol + Cystone (5 ml/kg) (Himalaya Lab., India) for 28 days. Group IV animals received 0.75% ethylene glycol in drinking water ad libitum along with aqueous extract of Crataeva magna 200mg/kg.  Group V animals received 0.75% ethylene glycol in drinking water ad libitum along with alcoholic extract of Crataeva magna 200mg/kg.

 

Urine and blood sampling

The crystalluria and stone formation was verified by different biochemical marker analysis of urine and serum.  The urine samples of the test animals in different groups were collected in their respective end day of the experiment.  The collected urine sample volume were measured followed by centrifugation at 3000 rpm for 10 minutes.  After centrifugation the urine samples were examined under light microscope (LAICA, DME Germany 400X) to ensure the presence of oxalate microcrystal followed by biochemical analysis (urine oxalate, calcium and uric acid).  The blood samples were collected from the animals under anaesthesia (ether) before sacrificing.  The collected blood samples were then centrifuged to obtain serum for the analysis of serum creatinine and serum calcium.

 

Table 1: Urine and serum parameters in control and experimental animals

Parameter

 

Group I

control

Group II

lithiasis induced

Group III

Cystone

Group IV

AqE

Group V

AlcE

Urine (mg/dl)

Oxalate

 

Calcium

 

Uric acid

0.55 ± 0.04

3.15 ± 0.22

0.73 ± 0.06

5.03 ± 0.24

4.27 ± 0.31

2.2 ± 0.12

2.31± 0.21

3.31± 0.31

0.71± 0.04

4.55 ± 0.11

4.12 ± 0.14

1.88 ± 0.19

4.34 ± 0.41

3.43 ± 0.31

1.78 ± 0.19

  Serum (mg/dl)

Creatinine

 

Calcium

 

8.40 ± 0.42

5.11± 0.31

9.22± 0.40

3.31± 0.21

8.12± 0.21

3.11 ±0.24

8.92± 0.61

2.81± 0.11

8.58± 0.31

3.01±  0.15

 

Statistical analysis:

Results were indicated in terms of Mean ± SEM.  Statistical significance of data were determined by One way-ANOVA followed by comparison between different groups using ‘Tukey Kramer’ multiple comparison test.  Differences between the data were considered significant at P<0.05.

 

RESULTS AND DISCUSSION:

In the present study, male rats were selected to induce urolithiasis because the urinary system of male rats resembles that of humans5  and also earlier studies have shown that the amount of stone deposition in female rats was significantly less6.  Urinary supersaturation with respect to stone-forming constituents is generally considered to be one of the causative factors in lithiasis.  Evidence in previous studies indicated that in response to 14 day period of ethylene glycol (0.75%) administration, young male albino rats form renal calculi composed mainly of calcium oxalate7.  Urinary volume is markedly increased in experimental groups. The increased urine volume in all the drug-treated groups might be due to the diuretic effect of the drug.  Similar results are also observed when lupeol is used as an antiurolithic agent8,9.

 

The results of urine and serum biochemistry showed significant reduction in urine calcium, uric acid and oxalate level, serum calcium with significant elevation in urine volume output, the markers previously reported which affirmed potent antiurolithiatic activity7. The results of serum and urine biochemistry is indicated in Table 1 comprises, the urine and serum biochemistry data of  Ethylene glycol (0.75%) induced lithiasis model data.  There was a significant increase in the urine output of Ethlene glycol treated rats (P < 0.001) in the 7th day of Ethylene glycol treatment.  Treatment with Aqueous and Alcoholic extracts reduced the oxalate excretion significantly.  A maximum oxalate excretion was observed with EG- treated rats on the 28th day.  Calcium excretion was increased in Ethylene glycol treated rats on day 28.  Administration of Crataeva magna decreased the calcium excretion significantly.

 

CONCLUSION:

Kidney stone disease has afflicted humankind since antiquity and can persist, with serious medical consequences, throughout a patient’s lifetime.  In addition, the incidence of kidney stones has been increased in most societies in the last five decades, especially in association with economic development.  According to our results, leaves of Crataeva magna was effective for prevention and treatment of Calcium Oxalate kidney stone in rats.  A dose of 200mg/kg of the plant significantly decreased the number and size of Calcium oxalate deposits in different segments of the renal tubules.  A higher dose of the plant leaves had also preventive and therapeutic effects on calcium oxalate kidney stone.  Further studies to determine the same effects on human beings are recommended.

 

REFERENCES:

1.       Smith, C.L. and D.R.P. Guay, 1992. Nephrolithiasis.  In: Pharmacotherapy: A Pathophysiologic Approach, DiPiro, J.T., R.L. Talbert, P.E. Hayes, G.C. Yee, G.R. Matzke and L.M. Posey (Eds.), 2nd Edn., Elsevier, New York, ISBN: 0838579760, pp: 720.

2.       Tiselius HG, Etiology and investigation of stone disease, Curriculum in Urology, Eur Urol, 1998, 2(1), 1-7.

3.       Acckart KSJ, Schroder FH.  Effect of extra corporeal shock wave lithotripsy (ESWL) on renal tissue.  Urol Res 1989; 17:   3-7.

4.       Prabhat Das, Ranjan Sethi, S. Mekap, S. Pani., Phytochemical and Pharmacological Screening of the Plant Crataeva magna against Alloxan Induced Diabetes in Rats.  J. Pharm. Sci and Res. Vol. 2 (4), 2010, 257-263.

5.       Vermeulen, C.W., 1962.  Experiments on causation of urinary calculi. In: Essays in Experimental Biology.  University of Chicago Press, Chicago, pp. 253-269.

6.       Prasad, K.V.S.R.G., Bharathi, K., Srinivasan, K.K., 1993.  Evaluation of Musa (Paradisiaca Linn Cultivar)- “Puttubale” stem juice for antilithiatic activity in albino rats.  Indian Journal of Physiology and Pharmacology 37, 337-341.

7.       Selvam, P., Kalaiselvi, P., Govindaraj, A., Murugan, V.B., Sathiskumar, A.S., 2001.  Effect of A. lanata leaf extract and Vediuppu chunnam; on the urinary risk factors of calcium oxalate urolithaisis during experimental hyperoxaluria.  Pharmacological Research 43, 89-93.

8.       Anand R, Patnaik GK, Kulshrestha DK, Dhawan BN, Antiurolithiatic and diuretic activity of lupeol. Proc 24th and 25th Pharmacol Soc Conf 1991; A-10.

9.       Malini MM, Baskar R, Varalakshmi P.  Effect of lupeol, a pentacyclic triterpene on urinary enzyme in hyperoxaluric rats, Jpn J Med Sci Biol 1995; 48: 211-20.

 

Received on 17.06.2011

Modified on 29.06.2011

Accepted on 08.07.2011           

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Research J. Science and Tech.  3(5): Sept.-Oct. 2011: 295-297