Depleted uranium (DU) has been widely used in both civilian and military activities and contributes to health problems. This study was undertaken to evaluate the protective role of zinc against acute toxicity of DU. Sprague Dawley rats were injected with DU (10 mg/kg, i.p.) to create a toxicity model (DU group). Before and after the injection of DU, zinc sulphate (10 mg/kg, i.p.) was administered once a day for 2 days. The survival rates at 30 days post DU administration and the effects of zinc at 4 days post DU administration were evaluated. Our data indicate that zinc has obvious protective effects, especially pre-treatment with zinc. Rats pre-treated with zinc had significantly higher survival rates than rats in the DU group, with 60.03% more surviving. In addition, at 4 days post DU administration, the former had lower kidney uranium content, insignificant renal tubular epithelial cell necrosis and less transparent tubes. Meanwhile, blood urea nitrogen, creatinine and urine N-acethyl-β-d-glucosaminidase concentrations were significantly decreased; the gene expression levels of metallothionein (MT) in kidney tissues were significantly increased; and catalase levels were increased and malondialdehyde levels were decreased. In conclusion, pre-treatment with zinc significantly alleviated acute toxicity of DU, and the mechanism appeared to be related to the induction of MT synthesis and enhancement of the antioxidant function.