|Year : 2011 | Volume
| Issue : 4 | Page : 466-468
Effect of Unex on ethylene glycol-induced urolithiasis in rats
Elias Edwin Jarald1, Pankaj Kushwah1, Sheeja Edwin1, Suhail Asghar2, Showkat Ahmad Patni2
1 TIFAC CORE in Green Pharmacy, B. R. Nahata College of Pharmacy, Mandsaur, Madhya Pradesh, India
2 Unijules Life Sciences Pvt. Ltd., Nagpur, Maharashtra, India
|Date of Submission||29-Dec-2010|
|Date of Decision||17-Apr-2011|
|Date of Acceptance||25-Apr-2011|
|Date of Web Publication||22-Jul-2011|
Elias Edwin Jarald
TIFAC CORE in Green Pharmacy, B. R. Nahata College of Pharmacy, Mandsaur, Madhya Pradesh
Source of Support: TIFAC-CORE, Conflict of Interest: None
This study was aimed to evaluate the effectiveness of the Unex capsule on albino rats as a preventive agent against the development of kidney stones. The Unex capsule is a marketed product of Unijules Life Sciences, Nagpur, containing the extracts of Boerhaavia diffusa and Tribulus terrestris. Activity of Unex was studied using the ethylene glycol-induced urolithiasis model. Standard drug used was Cystone. Several parameters were used including urinary volume, urine pH, urine analysis, and serum analysis to assess the activity. The results indicated that the administration of Unex to rats with ethylene glycol-induced lithiasis significantly reduced and prevented the growth of urinary stones (P < 0.01). Also, the treatment of lithiasis-induced rats by Unex restored all the elevated biochemical parameters (creatinine, uric acid, and blood urea nitrogen), restored the urine pH to normal, and increased the urine volume significantly (P < 0.01) when compared to the model control drug. This study supports the usage of Unex in urolithiasis and the utility could further be confirmed in other animal models.
Keywords: Boerhaavia diffusa, ethylene glycol, Tribulus terrestris, unex, urolithiasis
|How to cite this article:|
Jarald EE, Kushwah P, Edwin S, Asghar S, Patni SA. Effect of Unex on ethylene glycol-induced urolithiasis in rats. Indian J Pharmacol 2011;43:466-8
|How to cite this URL:|
Jarald EE, Kushwah P, Edwin S, Asghar S, Patni SA. Effect of Unex on ethylene glycol-induced urolithiasis in rats. Indian J Pharmacol [serial online] 2011 [cited 2019 Nov 19];43:466-8. Available from: http://www.ijp-online.com/text.asp?2011/43/4/466/83124
| » Introduction|| |
Urinary stone disease has affected humankind since antiquity and can persist, with serious medical consequences, throughout the patient's lifetime. In addition, the incidence of kidney stones has been increased in western societies in the last five decades, in association with economic development. Most calculi in the urinary system arise from a common component of urine, calcium oxalate (CaO), representing up to 80% of analyzed stones. ,, This may cause obstruction, hydronephrosis, infection, and hemorrhage in the urinary tract system. Surgical operation, lithotripsy, and local calculus disruption using high-power laser are widely used to remove the calculi.  Many remedies have been employed since ages to treat renal stones and most of them were from plants and proved to be useful. ,, The present day medical management of nephrolithiasis is either costly or not without side effects.  The current study was aimed to evaluate the effectiveness of the Unex capsule on albino rats as a preventive agent against the development of kidney stones. The Unex capsule (Herbajules Tricare in Malaysia) is a marketed product of Unijules Life Sciences, Nagpur, containing the extracts of Boerhaavia diffusa and Tribulus terrestris.
| » Materials and Methods|| |
Procurement of Materials
The product Unex containing the extracts of B. diffusa and T. terrestris was provided by the Unijules Life science Ltd. and Associated Companies, Nagpur. Cystone containing the extracts of Didymocarpus pedicellata, Saxifraga ligulata, Rubia cordifolia, Cyperus scariosus, Achyranthes aspera, Onosma bracteatum, Vernonia cinerea, Shilajeet (purified) and hajrul yahood bhasma was procured from Himalaya herbal health care, Bangalore. The urea kit, creatinine mono reagent test kit, and triglycerides test kit were procured from Central Store, BRNCOP.
Healthy adult male Wistar albino rats weighing between 150-200 g were selected for the antiurolithiatic activity. The animals were acclimatized to standard laboratory conditions (temperature: 25±2°C) and maintained on 12-h light:12-h dark cycle. They were provided with regular rat chow and drinking water ad libitum. The animal care and experimental protocols were approved by Institutional Animal Ethical Committee (IAEC) Reg no. (918/ac/05/CPCSEA).
Ethylene Glycol Induced Urolithiasis Model
Ethylene glycol-induced hyperoxaluria model was used to assess the antilithiatic activity in albino rats. Animals were divided into five groups containing six animals in each. Group I served as control and received regular rat food and drinking water ad libitum. Ethylene glycol (0.75%) in drinking water was fed to Groups II-V for induction of renal calculi for 28 days. Group III received standard antiurolithiatic drug, cystone (750 mg/kg body weight) from 15 th till 28 th day. Group IV and V received Unex at high dose (400 mg/kg) and low dose (200 mg/kg) (po) from 15 th till 28 th day. ,
Assessment of Antiurolithiatic Activity
Collection and analysis of urine
All the animals were kept in individual metabolic cages and urine samples of 24 h were collected on the 28 th day. Animals had free access to drinking water during the urine collection period. A drop of concentrated hydrochloric acid was added to the urine before being stored at 4°C. Urine was analyzed for calcium, phosphate, and oxalate content using the method of Bahuguna et al. 
After the experimental period, blood was collected from the retro-orbital under anesthetic condition and animals were sacrificed by cervical decapitation. Serum was separated by centrifugation at 10 000 × g for 10 min and analyzed for creatinine, uricacid, and urea nitrogen using the method of Atef and Attar. 
Animals were placed in separate metabolic cages for 24 h and total urinary volume was measured using the measuring cylinder and reported in ml. 
Uric acid crystals were found to deposit most frequently in the concentrated acid urine. Thus, the acidity of the urine was tested using the pH meter. 
Statistical evaluation was done using one-way analysis of variance (ANOVA) followed by the Bonferroni test. Statistical significant was set at P < 0.05. Results are presented as mean ± standard error of mean (SEM).
| » Results|| |
In this study, the administration of 0.75% (v/v) ethylene glycol aqueous solution to male Wistar albino rats produced hyperoxaluria. Oxalate, calcium, and phosphate excretion were grossly increased in the calculi-induced animals However, supplementation with Unex significantly lowered the elevated level of oxalate, calcium, and phosphate in urine when compared to the model control group [Table 1].
The level of serum uric acid, blood urea nitrogen (BUN), and creatinine were found to increase in the calculi-induced animal [Table 2]. In the case of Unex-treated groups, the treatment significantly (P < 0.001) lowered the elevated level of creatinine, uric acid, and BUN. Urine volumes were increased by Unex and the standard drug cystone compared to the model control group. Urinary pH was significantly increased in the animals treated with the 0.75% of ethylene glycol. Unex and the standard drug cystone significantly decreased the pH [Table 3]. No significant differences were observed in the activity of Unex between low dose and high dose.
| » Discussion|| |
Urinary supersaturation with respect to stone forming constituents is generally considered to be one of the causative factors in calculogenesis. Evidence in previous studies indicated that after 14 days period of ethylene glycol (0.75%, v/v) administration, renal calculi was formed in the young male albino rat composed mainly of calcium oxalate.  Stone formation in ethylene glycol-fed rats is caused by hyperoxaluria, which cause increased renal retention and excessive excretion of oxalate in urine.  In this study, oxalate and calcium excretion were increased in calculi-induced animals (Group II). An increase in urinary phosphate is also observed in calculi induced rats (Group II). Increased urinary phosphate excretion along with oxalate stress seems to provide an environment appropriate for stone formation by forming calcium phosphate crystals, which induce calcium oxalate deposition. Treatment with the Unex restored the phosphate level, thus reducing the risk of stone formation.  In urolithiasis, the glomerular filtration rate (GFR) decreases due to the obstruction to the outflow of urine by stones in the urinary system. Due to this, the waste products, particularly nitrogenous substances such as urea, creatinine, and uric acid get accumulated in blood. Also, increased lipid peroxidation and decreased levels of antioxidant potential have been reported in the kidneys of rats supplemented with a calculi producing diet. In this context, oxalate has been reported to induce lipid peroxidation and to cause renal tissue damage by reacting with polyunsaturated fatty acids in the cell membrane. In calculi-induced rats (Group II), marked renal damage was seen by the elevated serum levels of creatinine and uric acid, and BUN.  However, the curative treatment with product unex and cystone caused diuresis and hastened the process of dissolving the preformed stones and prevention of new stone formation in the urinary system. The diuretic effect of Unex was evident from urine volumes collected when compared to the model control group.
| » Conclusion|| |
In conclusion, the results indicated that the administration of Unex to rats with ethylene glycol-induced lithiasis reduced the growth of the urinary stone. The dose requirement of Unex to produce the activity is less than the standard drug used.
| » Acknowledgement|| |
The authors are thankful to the TIFAC-CORE for providing the grant necessary to carry out this research work.
| » References|| |
|1.||Ravindra VK, Navneet BG, Alagawadi KR, Rudraprabhu VS. Effect of Moringa oleifera Lam. root - wood on ethylene glycol induced urolithiasis in rats. J Ethnopharmacol 2006;105:306-4. |
|2.||Vyas BA, Vyas RB, Joshi SV, Santani DD. Antiurolithiatic activity of whole-plant hydroalcoholic extract of Pergularia daemia in rats. J Young Pharm 2011;3:36-40. |
|3.||Tania AV, Cristina DD, Ana PS, Maria TR, Antonio JL, Caden S. Evaluation of the antiurolithiatic activity of the extract of Costus spiralis Roscoe in rats. J Ethnopharmacol 1999;66:193-8. |
|4.||Hadjzadeh MA, Khoei A, Hadjzadeh Z, Parizady M. Ethanolic extract of Nigella sativa L seeds on ethylene glycol-induced kidney calculi in rats. Urol J 2007;4:86-90. |
|5.||Purnima A, Basavaraj CK, Vishwanathswamy AH. Antiurolithiatic and antioxidant activity of Mimusops elengi on ethylene glycol-induced urolithiasis in rats. Indian J Pharmacol 2010;42:380-3. |
|6.||Vargas SR, Perez GR, Perez GS, Zavala SM, Perez GC. Antiurolithiatic activity of Raphanus sativus aqueous extract on rats. J Ethnopharmacol 1999;68:335-8. |
|7.||Hossein H, Ali-Reza K, Zahra K, Vahideh M. Antiurolithiatic Activity of Pinus Eldarica Medw. Fruits Aqueous Extract in Rats. Urol J 2010;7:232-7. |
|8.||Bahuguna Y, Rawat MM, Juyal V, Gupta V. Antilithiatic effect of flower Jasmin auriculatum Vahl. Int J Green Pharm 2009;3:155-63. |
|9.||Khatib N, Dhaval G, Hashilkar N, Rajesh KJ. Antiurolithiatic potential of the fruit extracts of Carica papaya on ethylene glycol induced urolithiatic rats. J Pharm Res 2010;3:2772-5. |
|10.||Atef M, Attar A. Antilithiatic influence of spirulina on ethylene glycol-induced nephrolithiasis in male rats. Am J Biochem Biotechnol 2010;6:25-6. |
|11.||Anbu J, Suman S, Swaroop Kumar SL, Satheeshkumar R, Nithya S, Kannadhasan R. Antiurolithiatic Activity of Ethyl Acetate Root Extract of Ichnocarpus frutescens using Ethylene Glycol Induced Method in Rats. J Pharm Sci Res 2011;3:1182-9. |
|12.||Atmani F, Slimani Y, Minouni M, Hacht B. Prophylaxis of calcium stone by Hibiscus sabdariffa on experimentally induced nephrolithiasis in rats. BJU Int 2003;92:137-43. |
[Table 1], [Table 2], [Table 3]