Our aim was to investigate the protective role of ozone treatment against gentamicin-induced nephrotoxicity in an experimental rat model. In this study, a total of 30 rats were allocated in 5 groups (n=6 in each group). The control group (Group 1) received isotonic saline only, while Groups 2 and 3 received gentamicin at doses of 15 mg/kg/day and 50 mg/kg/day, respectively. In Group 4, intraperitoneal ozone treatment (1 mg/kg, 5% O-3-95% O-2) was performed after administration of gentamicin at a dose of 15 mg/kg/day. Group 5 underwent ozone treatment intraperitoneally following the application of gentamicin (50 mg/kg/day). Nephrotoxicity was formed by administration of glycerol.Serum levels of urea, creatinine, neutrophil-gelatinase-associated lipocalin (NGAL), lactate dehydrogenase (LDH), total antioxidant capacity (TAC) and protein carbonyl were measured, and kidneys were histopathologically examined after the sacrifice of animals on the 5th, day. Group 4 displayed more favorable outcomes regarding biochemical markers of oxidative stress such as NGAL, LDH, creatinine, urea, TAC and protein carbonyl. Similarly, histopathological alterations indicating gentamicin-induced nephrotoxicity such as hemorrhage, the presence of protein casts and epithelial injury in renal tubules were less evident in Groups 4 and 5 which received ozone treatment. To conclude, results of this experimental study demonstrated that ozone treatment might ameliorate biochemical disturbances and histopathological alterations linked with gentamicin-induced nephrotoxicity. However, furthertrials are warranted to document the actual therapeutic potential of ozone treatment in the clinical setting.