Reactive oxygen species (ROS) were shown to contribute to the cellular damage induced by ischemia-reperfusion. The purpose of this study was to investigate and compare the efficiency of melatonin and vitamin E in the reduction of injury induced by ROS in a rat model of renal ischemia-reperfusion. Twenty-four Wistar-albino rats were divided into four groups. Rats in the Sham group were given saline 1 mL/kg, intraperitoneally (ip) 72 h, 48 h, 24 h, and 30 min before the sham operation. Rats in ischemia-reperfusion (IR), IR+Melatonin, and IR+Vitamin E groups were given saline (1 mL/kg), melatonin (10 mg/kg), and vitamin E (100 mg/kg) ip, respectively, 72 h, 48 h, 24 h, and 30 min before the ischemia for 60 min, followed by reperfusion for 60 min. The blood samples and kidney tissues of the rats were taken under anesthesia. Ischemia-reperfusion significantly increased urea, creatinine, and malondialdehyde (MDA) levels, and decreased superoxide dismutase (SOD) and catalase (CAT) activities. Histopathological findings of the IR group confirmed that there was renal impairment by cast formation and tubular necrosis in the tubular epithelium. In the IR+Melatonin group, while MDA levels significantly decreased, SOD activities increased. In the IR+Melatonin group, the level of tubular necrosis and cast formation are significantly decreased than those seen in the ischemia-reperfusion group. Melatonin in particular was effective to reverse hot ischemia of kidney by its antioxidant effects. These results may indicate that melatonin pretreatment protects against functional, biochemical, and morphological damage better than vitamin E in renal ischemia-reperfusion injury.