Indian Journal of Experimental Biology Vol. 44, May 2006, pp. 416-418
Modification of biochemical parameters of gentamicin nephrotoxicity by Coenzyme Q10 and green tea in rats Aman Upaganlawar*, Mamta Farswan, Shivkumar Rathod & R Balaraman Pharmacy Department, Faculty of Technology and Engineering, Maharaja Sayajirao University of Baroda, Vadodara 390 001, India Received 26 December 2005; revised 27 February 2006 The present study was designed to investigate the possible potential protective role of coenzymeQ10 (CoQ10; 10mg/kg/day, ip) and/or green tea (GT; 25mg/kg/day, po) against gentamicin (GM) nephrotoxicity. Marked increase in the level of serum urea, creatinine and lipid peroxidation (LPO) content was found after administration of gentamicin (80mg/kg/day, ip) for eight days along with significant decrease in the antioxidant enzymes, superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT) as well as brush border enzymes (Na+/K+ ATPase, Mg+2ATPase and Ca2+ ATPase).Treatment with CoQ10 or green tea alone with GM showed significant decrease in serum urea, creatinine and tissue LPO content and significant increase in antioxidant and membrane bound enzymes. Combined treatment with CoQ10 and green tea was more effective in mitigating adverse effect of GM nephrotoxicity. The present work indicated that CoQ10 and green tea due to their antioxidant activity modified the biochemical changes occurred during gentamicin nephrotoxicity and thus had a potential protective effect. Keywords: Antioxidant, Coenzyme Q10, Gentamicin, Green tea, Nephrotoxicity, Oxidative stress
Gentamicin is an aminoglycoside antibiotic that has been widely used in the treatment of infections caused by Gram negative bacteria. Nephrotoxicity is the major side effect of aminoglycosides, accounting for 10-20% of all cases of renal failure1. These antibiotics are excreted by glomerular filtration and are partially reabsorbed by proximal tubules. The tubules also accumulate the antibiotics and hence are the primary sites for nephrotoxicity which is an important limiting factor in its clinical use2. Several in vivo and in vitro studies have demonstrated that reactive oxygen species including ________________ *Address for correspondence: Institute of Pharmaceutical Education & Research, Borgaon (Meghe), Wardha 442 001, India E-mail:
[email protected]
free radical, superoxide, hydroxyl radical anion and hydrogen peroxide are important mediators of tissue injury3. Oxygen free radicals have been implicated in several biological processes potentially important in glomerular disease, in renal ischemia, reperfusion injury4 and in various models of acute renal failure5. The protective effect of various therapeutic approaches aiming at reduction in formation of reactive oxygen species have already been reported in animal models6 .CoQ10 (Ubiquinone) is a fat soluble vitamine present in every cell of the body and serves as a coenzyme that is essential in the production of energy in cell and is a powerful free radical scavenger in lipid and mitochondrial membrane7. CoQ10 scavenges the free radicals generated in diabetic rats and in rhabdomyalosis8, 9. Green tea is a polyphenolic compound including epicatechin, epicatechin gallate, epigallocatechin gallate and proanthrocyanidine and is a potent antioxidant used in the treatment of several disorders like diabetes, atherosclerosis10, 11. No study has been reported about protective role of CoQ10 and green tea in nephrotoxicity and thus, an attempt has been made to carry out the possible protective role of CoQ10 and green tea alone and in combination in assessing the biochemical parameters during gentamicin induced nephrotoxicity. Sources of materials⎯CoQ10 and gentamicin were obtained as gift samples from Troikka laboratory, Ahmedabad, India and Alembic lab, Vadodara, India. Green tea (in powder form) was obtained from Indone Healthcare Ltd, Ahmedabad, India. All the chemicals of analytical grade required for biochemical estimation were purchased from HIMEDIA, Mumbai. Maintenance of animals⎯Male albino rats (Wistar strain) weighing 200-250g were maintained at 25°±2°C and kept in well ventilated animal house under natural photoperiodic condition in large polypropylene cages and were fed standard rat chow and water ad libitum. The animal experiment was approved by Animal Ethical Committee of the Institute. Animals were divided into different groups with six animals in each group. Group I (Gr I)-Served as control and received normal saline throughout experiment period; Group II (Gr II)⎯received gentamicin (80mg/kg/day, ip)1; Group III (Gr
SHORT COMMUNICATION
III)⎯received CoQ10 (10mg/kg/day, ip); Group IV (Gr IV)⎯received green tea (25mg/kg/day, po); Group V (Gr V)⎯received gentamicin concomitantly with CoQ10; Group VI (Gr VI)⎯received gentamicin concomitantly with green tea; Group VII (Gr VII)⎯Received gentamicin concomitantly with CoQ10 and green tea. The treatment periods for all these groups were eight days. Biochemical assay⎯After respective treatments of the animals for 8 days, blood was collected from orbital sinus of rat under ether anesthesia and centrifuged using the table top centrifuge (REMI centrifuge) at 2000 rpm so as to get the serum. Level of urea and creatinine was estimated using Span diagnostic kit on Chemical analyzer (Microlab 3000) for assessment of nephrotoxicity. Kidney was removed, homogenized and centrifuged at 10,000 rpm at 0°C for 20 min. The supernatant was used for the estimation of different antioxidant levels by calorimetric method using ultraviolet double beam spectrophotometer (Hitachi U-2000). Lipid peroxidation estimated by 12 thiobarbituric acid (TBA) methods . Antioxidant enzymes such as superoxide dismutase (SOD)13, catalase (CAT)14 and glutathione reductase (GSH)15 were measured as described earlier. The sediment from homogenate was used for estimation of Na+/K+ ATPase, Mg+2ATPase and Ca2+ ATPase by the method of Bonting16, Hijesken17 and Ohinish18, respectively. Statistical analysis⎯One way analysis of variance (ANOVA) followed by Dunnett’s t test was carried out and P
