Gentamicin is the most common antibiotic in group of aminoglycosides. However, due to its side effects, there are some limitations in the use of gentamicin. In this study, 30 rats were divided to 6 equal groups randomly. Then, the drugs were injected by the route of I.M, once daily, for 10 days as followed. Group1: gentamicin 80 mg/kg. Group2: normal saline. Group3, 4,5 gentamicin 80 mg/kg plus vitamin B6 in doses 100,150,200 mg/kg respectively. Group6: vitamin B6 200mg/kg. At the end of experiment, animals were euthanized and serum creatinine and blood urea nitrogen (B.U.N) were measured. Statistical analysis showed that according to the concentration of creatinine, there is no any significant difference between group 2 and 5.While significant difference has seen between each of group 2 and 5 with other groups and also between group1 and other groups. On the other hand, based on the B.U.N values, there is no any significant difference between group2 and 5 and 6.While significant difference has seen between each of group 2,5 and 6 with group 1,3 and 4 and also between group 1 and all other groups. Therefore, the results showed that vitamin B6 in high doses can decrease renal toxicity of gentamicin. This effect is probably due to decreasing gentamicin level in plasma and kidney by connecting piridoxal-5-phosphate to gentamicin. In addition, vitamin B6 can increase regeneration in injured proximal convoluted tubules.