EFFECT OF ETHANOLIC EXTRACT OF ISAPGOL AGAINST DIET INDUCED UROLITHIASIS IN RATS.
*SOUMYA G., SRINIVAS S., NAVEEN KUMAR M., KRISHNA MOHAN C.
Department of Pharmacology,
St. John College of Pharmacy, Warangal,
Andhra Pradesh, India-5067371.
Kidney stone formation or urolithiasis is a complex process that is a consequence of an imbalance between promoters and inhibitors in the kidneys. Even though the technological developments in the present medical practice the formation and growth of renal calculi continues to afflict humankind. Though various kinds of stone have been identified, calcium stones are the most common in human as well as rats. The rat experimental models of Calcium oxalate urolithiasis, induced by ethylene glycol alone, or in combination with other drugs such as ammonium chloride are often used to study the pathogenesis of kidney crystal deposition. The urolithiasis was induced by administration of gentamycin and calculi producing diet (5% ammonium oxalate in standard rat pellet feed) for 28 days to rats’ results in hyperoxaluria, Calcium oxalate crystalluria, and occasional deposition of CaOx crystals in the kidney.
Reference Id: PHARMATUTOR-ART-1443
MATERIALS AND METHODS
The plant of Isapgolwere collected from the surroundings of Chittor Disrict, Andra Pradesh and authenticated by Dr. K. Madhava chetty Department of Botany, Sri Venketeswara Univerisity.
Preparation of Methanolicextract
The plant of Isapgol were cleaned and chopped into small pieces and dried under shade. Thecoarse powder by mechanical grinding. The powdered material 100 g was subjected to continue hot extraction in soxhlet apparatus at a temperature of (60- 700 c) by using Methanol (95% v/v). After complete extraction, the extract was dried. The yield was about 5% w/w and it was stored at 4oC in desiccator. The extract was diluted with 5ml of dimethylsulphoxide as suspending agent for oral administration to animals.
Selection of the dose
The LD50 value of methanolic extracts Isapgol of plant has been reported. Hence, one 10th of LD50 dose was taken as a therapeutic dose for determination of antiurolithiatic effect.
Male albino rats of Wistar rats weighing between 150-200g were used. The animals were fed with commercial rat feed pellets and were given water ad libitum. The animals were acclimatized and maintained at 240C and 12h/12h light dark cycle and were kept in polypropylene cages in a well-ventilated room under hygienic conditions throughout the study.
Gentamycine was procured from Ranbaxy, India. All other chemicals used are of analytical grade and obtained from S.D. Fine Ltd., India.
Antiurolithiatic studies- calcium oxalate stones
Hyperoxaluria and calcium oxalate deposition in the kidney was induced using gentamycine (40 mg/kg/s.c) and calculi producing diet (CPD). The latter was made from powdered standard rat pellet feed (Hindustan liver Ltd., Bombay, India) mixed with ammonium oxalate (5%), then made into pellets and dried.
Pharmacological screening for anti urolithiatic activity
Animals were divided into five groups, each containing six animals. Group I served as normal control and maintained on regular laboratory diet and water ad libitum. Group II to V were fed with gentamycine and calculi producing diet (5% ammonium oxalate in standard rat pellet feed) in standard rat pellet for induction of renal calculi till1-28th day. Group III received standard antiurolithiatic drug cystone (750 mg/kg body weight) from 15th to 28th day. Group IV served as curative regimen, received methanolic extract of the plant of Isapgol at a dose of 400 mg/kg, body weight from 15th day to 28th day respectively. Groups V served as preventive regimen received methanolic extract of the plant of Isapgol at a dose of 400 mg/kg body weight from 1st day to 28th day respectively. Both the extracts were administered once daily by oral route.
Antiurolithiatic activity: gentamicine and CPD induced urolithiatic model
Assessment of urinary parameters
The rats were hydrated with 5ml of distilled water (p.o) and placed in separate metabolic cages and 24hr urine samples were collected. Urine samples were collected from Normal, Curative and Preventive groups after 28 days interval and at the end of the experimental period.The collected urine was analyzed for calcium and using standard methods. The volume of urine collected from all groups was recorded. Further microscopy of the urine was performed.
Kidney homogenate analysis
At the end of experimental period, the rats were sacrified and kidneys were carefully removed, washed in ice cold 0.15M KCl and their weights were recorded. One kidney was homogenized in 10% Hcl. The homogenate was centrifuged at 2500 rpm and supernatant was used for estimating calcium and phosphorus.
Results are expressed as mean ± SEM. Statistical analysis was carried out using one-way ANOVA followed by Bonferroni’s Multiple Comparison test. Differences between the data were considered significant at P<0.05.
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