Drug-induced nephrotoxicity is an important cause of renal failure in dogs. Aminoglycoside antibiotics, such as gentamicin, can produce nephrotoxicity in dogs, due to in part to an imbalance of pro- and antioxidants (oxidative stress). Silymarin (the mixture of flavonolignans extracted from Silybum marianum) has potentially beneficial antioxidant properties. A control group (saline, group 1, n = 5) was compared with dogs that were administrated gentamicin by intramuscular injection, at dosage of 20 mg/kg, once daily for 9 days (groups 2-5, n = 5 per group). The effects of vitamin E (group 3) and silymarin (group 4) alone and in combination (group 5) were compared for induced nephrotoxicity. Renal function was assessed using serum biochemical markers (creatinine and urea). Malondialdehyde (MDA) concentration were measured as a marker of lipid peroxidation. The activity of total serum antioxidants (TSAO) was assessed as a marker of antioxidant defences. Serum creatinine and urea concentrations were increased significantly and TSAO was decreased significantly in group 2 compared with group 1. Serum creatinine concentrations but not urea concentrations were significantly lower in groups 3 and 4 than in group 2 (P = 0.001). Serum MDA concentrations was significantly different between groups 2 and 3 (P = 0.01), 2 and 4 (P < 0.001) and 4 and 5 (P = 0.01). TSAO activity was significantly in group 4 (silymarin) than in group 2 (P = 0.002). Silymarin and vitamin E decreased gentamicin-induced nephrotoxicity in dogs.