Although methotrexate (MTX) is an effective immunosuppressive and anti-cancer agent, it is associated with side effects, including nephrotoxicity. Capsaicin, a component of hot chilli peppers, induces rapid desensitization of TRPV1 pain receptors and therefore has uses in pain treatment. Capsaicin also has anti-cancer activity, including anti-inflammatory properties. Thus, capsaicin may have potential in preventing MTX-induced nephrotoxicity. The purpose of this research work was to observe protective effects of capsaicin towards renal toxicity caused by methotrexate and mechanisms responsible for these effects. As expected, capsaicin had nephroprotective effects in MTX-intoxicated rats. Serum creatinine urea, nitric oxide (NO) and renal malondialdehyde (MDA) levels decreased significantly, with a concurrent increase in superoxide dismutase (SOD) and renal glutathione peroxidase (GPx) activities as compared to rats that had been untreated with nephrotoxic. Biochemical analyses confirmed the protective effects of capsaicin. We conclude that capsaicin provides protection against MTX-nephrotoxicity in rats via anti-inflammatory and antioxidant activities.