Nanoparticles of zinc oxide (ZnO NPs) are applied in many fields nowadays. Consequently, concerns have been raised about its potential harmful effects. The present study focuses on its toxic effect on podocytes and rats. In vitro study, podocytes were treated with different concentrations of ZnO NPs (10, 50 and 100 μg/ml), the viability of cells was decreased as time prolonged according to MTT assay. Meantime, flow cytometry analysis indicated that ZnO NPs induced intracellular accumulation of reactive oxygen species (ROS) and apoptosis. The measurement of superoxide dismutase (SOD) and malondialdehyde (MDA) showed that ZnO NPs decreased SOD level and increased MDA level. Interestingly, pretreatment with N-mercaptopropionyl-glycine, known as a type of ROS scavenger, could inhibit podocyte apoptosis induced by ZnO NPs. Meantime, a loss of nephrin can be detected, which may result in a direct damage to slit diaphragms. In vivo study, adult male Wistar rats were administrated with 3mg/kg/day ZnO NPs for 5 days, body weight and kidney index were significantly reduced. In addition, ZnO NPs decreased the activity of catalase and SOD in kidney cortex in vivo. It could be concluded that ZnO NPs present toxic effect on podocytes and Wistar rats, which was related with oxidative stress.