The influence of acute iron and ethanol load on lipid peroxidation and antioxidative defense systems in rat testes and the modification after supplemented with Vitamin E was investigated. Acute iron and ethanol load was achieved by i.p. injection of iron-dextran (500 mg/kg) and ethanol (50 mmol/kg). After 18 h, a significant increase in testis total iron content was induced. Compared with control, total testis iron content was 6.8-fold higher in iron-treated rats and 9.1-fold in iron and ethanol treated rats. As the content of iron increasing, the endogenous lipid peroxidation evaluated as 2-thiobarbituric acid-rective substances (TBARS) increased apparently, and the content of lipid-soluble antioxidants alpha-tocopherol decreased. The supplement of an antioxidant, alpha-tocopherol, protected against lipid peroxidation. Iron and ethanol treatment did not affect the activity of superoxide dismutase, catalase and glutathione peroxidase. The results indicate that acute iron load causes iron accumulation in rat testes and ethanol increases the same accumulation. Iron played pivotal paracrine roles on ethanolinduced injure rat testis. The supplement of Vitamin E can protect against this damage.