The goal of the present work was to investigate the protective effect of pumpkin seed oil (PSO) against lead acetate toxicity in male mice. lead acetate (20 mg/kg b.w.) was administered orally to mice once a day for 30 successive days. Whereas, PSO was administered to the mice orally at 1.5 mL/kg b.w. once a day for 30 successive days concurrently with treatment of lead acetate. The studied parameters were relative organs weights, DNA damage evaluation using comet assay in liver and kidney cells and micronucleus test in bone marrow and biochemical assessment of the liver and kidney function. Results showed that, lead acetate caused DNA damage in tested cells and significant increase in the levels of biochemical measurements. In contrast, PSO administration plus lead acetate effectively alleviated DNA damage in tested cells and improved the biochemical alterations. It can be concluded that PSO may has a protective role against lead acetate toxicity in male mice.