Methotrexate, a folic acid antagonist, is a chemotherapeutic agent frequently employed in treating certain forms of cancer. Nephrotoxicity caused by high-dose Methotrexate is a medical emergency due to the delayed renal excretion of Methotrexate. Chromium is a trace element found in certain foods and the environment. The primary function of chromium is to regulate glucose levels, which is used as a supplement by people with type 2 diabetes Mellitus. This study aimed to evaluate the renoprotective effect of chromium against Methotrexate-induced Nephrotoxicity in rats. Thirty-two Wistar rats were allocated into four groups and treated as Group-1 (Control group): The rat was administered distilled water orally as a vehicle for eight consecutive days. Group-2 (Induction group): Methotrexate was injected in rats at a single dose of 20 mg/kg intraperitoneally on the first day. On the following day, the rats received distilled water for seven days. Group-3 (Chromium 2mg): methotrexate injected Rats at a dose of 20 mg/kg intraperitoneally, as a single dose on the first day. On the following day, the rats received Chromium picolinate at a dose (2mg/kg) orally by oral gavage for seven days. Group-4 (Chromium 4mg): Methotrexate injected in rats at a dose of 20 mg/kg intraperitoneally, as a single dose on the first day. On the following day, the rats receive Chromium picolinate at a dose (4mg/kg) orally by oral gavage for seven days. The Results of our study indicated that injection (20mg/kg) of Methotrexate caused a significant increase in creatinine and blood urea nitrogen, malondialdehyde, and a significant reduction in superoxide dismutase-1. Interestingly, treatment with chromium picolinate (2mg/kg) and (4mg/kg) showed significant drops in creatinine and blood urea nitrogen and malondialdehyde and a significant elevation in superoxide dismutase-1   compared to Methotrexate-treated rats. In conclusion, the current investigation revealed that the administration of chromium to rats has a beneficial impact on reducing Nephrotoxicity generated by Methotrexate. This is achievd by enhancing antioxidant defenses of the kidneys.