Wistar rats were used to study the protective effect of zinc-induced metallothionein (MT) synthesis on gentamicin nephrotoxicity. We found that s.c. pre-injection of ZnSO4 (Zn, 10 mg/kg/day) for 5 days could ameliorate proximal tubular necrosis and acute renal failure caused by an 8-day s.c. injection of gentamicin (100 mg/kg/day), while preinjection of saline instead of zinc or zinc and gentamicin together could not. In the zinc-pretreated rats (n = 6), renal cortical metallothionein level was significantly higher than that of normal (n = 8, p < 0.001) and the saline controls (n = 6, p < 0.001). Since MT is a scavenger of hydroxyl radical, it is proposed that hydroxyl radical plays a role in the pathogenesis of gentamicin nephrotoxicity and that preinjection of zinc could ameliorate gentamicin nephrotoxicity via the induction of renal cortical MT synthesis.