Antioxidant properties of repaglinide and its protections against cyclosporine A-induced renal tubular injury

Document Type : Original Article


1 Department of Pharmacology, City College, Wuhan University of Science and Technology, Wuhan, China

2 Department of Laboratory, Zhongnan Hospital of Wuhan University, Wuhan, China


Objective(s): Repaglinide (RG) is an antihyperglycemic agent used for the treatment of non-insulin-dependent diabetes mellitus. It has a good safety and efficacy profile in diabetic patients with complications in renal impairment and is an appropriate treatment choice, even for individuals with more severe degrees of renal malfunctions. The aim of the present study was to examine the protective effect of RG on cyclosporine A (CsA)-induced rat renal impairment and to evaluate the antioxidant mechanisms by which RG exerts its protective actions.
Materials and Methods: Fifty male Sprague-Dawley rats weighing 250–300 g were randomly divided into five groups: administrations of olive oil (control, PO), RG (0.4 mg/kg, PO), CsA (30 mg/kg in olive oil, SC), RG (0.2 or 0.4 mg/kg, PO) plus CsA (30 mg/kg in olive oil SC) every day for 15 days.
Results: SC administration of CsA (30 mg/kg) to rats produced marked elevations in the levels of renal impairment parameters such as urinary protein, N-acetyl-beta-D-glucosaminidase (NAG), serum creatinine (SCr),and blood urea nitrogen (BUN).  It also caused histologic injury to the kidneys. Oral administration of RG (0.2 and 0.4 mg/kg) markedly decreased all the aforementioned changes. In addition, CsA caused increases in the levels of malondialdehyde (MDA) and decreases in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione reductase (GSR), glutathione-S-transferase (GST), and glutathione in kidney homogenate, which were reversed significantly by both doses of RG.
Conclusion: The findings of our study indicate that RG may play an important role in protecting the kidney from oxidative insult.


1. Polonsky KS, Given BD, Hirsch LJ, Tillil H, Shapiro ET, Beebe C, et al. Abnormal patterns of insulin secretion in non-insulindependent diabetes. N Engl J Med 1988; 318:1231–1239.
2. Hasslacher C. Safety and efficacy of repaglinide in type 2 diabetic patients with and without impaired renal function. Diabetes Care 2003; 26:886–891.
3. Yale JF. Oral antihyperglycemic agents and renal disease: new agents, new concepts. J Am Soc Nephrol 2005; 16:7–10.
4. Gumieniczek A.Oxidative stress in kidney and liver of alloxan-induced diabetic rabbits: effect of repaglinide. Acta Diabetol 2005; 42:75–81.
5. Tankova T, Koev D, Dakovska L, Kirilov G. The effect of repaglinide on insulin secretion and oxidative stress in type2 diabetic patients. Diabetes Res Clin Pract 2003; 59:43–49.
6. Haugen E, Nath KA. The involvement of oxidative stress in the progression of renal injury. Blood Purif 1999; 17:58–65.
7. Deponte M. Glutathione catalysis and the reaction mechanisms of glutathione-dependent enzymes. Biochim Biophys Acta 2013; 1830:3217–3266.
8. Ponticelli C. Cyclosporine: from renal transplan-tation to autoimmune diseases. Ann NY Acad Sci 2005; 1051: 551–558.
9. Busauschina A, Schnuelle P, Van der Woude FJ. Cyclosporine nephrotoxicity. Transplant Proc 2004; 36:229S–233S.
10. O'Connell S, Tuite N, Slattery C, Ryan MP, McMorrow T. Cyclosporine A induced oxidative stress in human renal mesangial cells: a role for ERK 1/2 MAPK signaling. Toxicol Sci 2012; 1261:101–113.
11. Ateşşahin A, Çeribaı OA, Yılmaz S. Lycopene, a carotenoid, attenuates cyclosporine-induced renal dysfunction and oxidative stress in rats. Basic Clin Pharmacol Toxicol 2007; 100:372–376.
12. Hagar HH, Eman EE, Maha A: Taurine attenuates hypertension and renal dysfunction induced by cyclosporine A in rats. Clin Exp Pharmacol Physiol 2006; 33:189–196.
13. Salant DJ, Cybulsky AV. Experimental glomerulo-nephritis. Methods Enzymol 1988; 162:421–461.
14. Price RJ: Urinary N-acetyl-β-D- glucosaminidase (NAG) as an indicator of renal disease. Curr Probl Clin Biochem 1979; 9:150–163.
15. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351–358.
16. Misra HP, Fridovich I. The role of superoxide anion in the autooxidation of epinephrine and a simple assay for superoxide-dismutase. J Biol Chem 1972; 247:3170–3175.
17. Beutler E, Durom O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med 1963; 61:882–888.
18. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 1974; 249:7130–7139.
19. Flohe L, Gunzler WA. Assays of glutathione peroxidase. Methods Enzymol 1984; 105:114–121.
20. Smith IK, Vierheller TL, Thorne CA. Rassay of glutathione reductase in crude tissue homogenates using 5,5'-dithiobis (2-nitrobenzoic acid). Anal Biochem 1988; 175:408–413.
21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent. J Biol Chem 1951; 193:265–275.
22. Gao H, Zhou YW. Inhibitory effect of picroside II on hepatocyte apoptosis. Acta Pharmacol Sin 2005; 26:729–736.
23. Marnett LJ. Lipid peroxidation-DNA damage by malondialdehyde. Mutat Res 1999; 424:83–95.
24. Del RD, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutr Metab Cardiovasc Dis 2005; 15: 316–328.
25. Kim J, Jung KJ, Park KM. Reactive oxygen species differently regulate renal tubular epithelial and interstitial cell proliferation after ischemia and reperfusion injury. Am J Physiol Renal Physiol 2010; 298:F1118–1129.
26. Khan RA, Khan MR, Sahreen S. Evaluation of Launaea procumbens use in renal disorders: a rat model. J Ethnopharmacol 2010; 128:452–461.
27. Aikemu A, Yusup A, Umar A, Berké B, Moore N, Upur H. The impact of the Uighur medicine abnormal savda munziq on antitumor and antioxidant activity in a S180 and Ehrlich ascites carcinoma mouse tumor model. Pharmacogn Mag 2012; 8:141–148.
28. Catalá A. Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chem Phys Lipids 2009; 157:1–11.
29. Yadav P, Sarkar S, Bhatnagar D. Action of Capparis deciduas against alloxan-induced oxidative stress and diabetes in rat tissues. Pharmacol Res 1997; 36:221–228.
30. Gumieniczek A. Effects of repaglinide on oxidative stress in tissues of diabetic rabbits. Diab Res Clin Pract 2005; 68:89-95.
31. Maritim AC, Sanders RA, Watkins JB. Effects of α-lipoic acid on biomarkers of oxidative stress in streptozotocin-induced diabetic rats. J Nutr Biochem 2003; 14:288–294.
32. Raza H. Dual localization of glutathione S-transferase in the cytosol and mitochondria: implications in oxidative stress, toxicity and disease. FEBS J 2011; 278:4243–4251.
33. Brezniceanu ML, Lau CJ, Godin N, Chénier I, Duclos A, Ethier J, et al. Reactive oxygen species promote caspase-12 expression and tubular apoptosis in diabetic nephropathy. J Am Soc Nephrol 2010; 21:943–994.
34. Habib SL. Alterations in tubular epithelial cells in diabetic nephropathy. J Nephrol 2013; 26:865–869.
35. Gilbert RE, Cooper ME. The tubulointerstitium in progressive diabetic kidney disease: more than an aftermath of glomerular injury? Kidney Int 1999; 56:1627–1637.
36. Backman JT, Kajosaari LI, Niemi M, Nevvonen PJ. Cyclosporine A increases plasma concentrations and effects of repaglinide. Am J Transplant 2006; 6:2221–2222.