Background and purpose: Ifosfamide nephrotoxicity is a serious adverse effect for children undergoing cancer chemotherapy. Our recent in vitro studies have shown that the antioxidant N-acetylcysteine (NAC), which is used extensively as an antidote for paracetamol (acetaminophen) poisoning in children, protects renal tubular cells from ifosfamide-induced toxicity at a clinically relevant concentration. To further validate this observation, an animal model of ifosfamide-induced nephrotoxicity was used to determine the protective effect of NAC. Experimental approach: Male Wistar albino rats were injected intraperitoneally with saline, ifosfamide (50 or 80 mg kg À1 daily for 5 days), NAC (1.2 g kg À1 daily for 6 days) or ifosfamide þ NAC (for 6 days). Twenty-four hours after the last injection, rats were killed and serum and urine were collected for biochemical analysis. Kidney tissues were obtained for analysis of glutathione, glutathione S-transferase and lipid peroxide levels as well as histology analysis. Key results: NAC markedly reduces the severity of renal dysfunction induced by ifosfamide with a significant decrease in elevations of serum creatinine (57.8 ± 2.3 vs 45.25 ± 2.1 mmol l À1 ) as well as a reduced elevation of b 2 -microglobulin excretion (25.44±3.3 vs 8.83±1.3 nmol l À1 ) and magnesium excretion (19.5±1.5 vs 11.16±1.5 mmol l À1 ). Moreover, NAC significantly improved the ifosfamide-induced glutathione depletion and the decrease of glutathione S-transferase activity, lowered the elevation of lipid peroxides and prevented typical morphological damages in renal tubules and glomeruli. Conclusions and implications: Our results suggest a potential therapeutic role for NAC in paediatric patients in preventing ifosfamide nephrotoxicity.