The aim of this study was to investigate the effects of P/S ratio of fatty acid and antioxidant (vitamin E, selenium) supplements on the serum lipid levels and hepatic antioxidant enzyme activity in rats. Female 16-week-old Sprague-Dawley rats were fed 6 different experimental diets for 4 weeks. While the peroxidizability index (PI) levels of fatty acids in the experimental diets were fixed at 81.22, the levels of P/S ratio of fatty acids were formulated at 0.38, 1.00, 4.81 (LP, MP, HP). These diets were supplemented with vitamin E (1,000 mg/kg diet) and selenium (2.5 mg/kg diet) (LP-S, MP-S, HP-S). This study showed that the serum concentrations of total-cholesterol and HDL-C increased with the increasing of the P/S ratio in the diet (p <0.05). Antioxidant supplementation significantly lowered the concentrations of triglyceride (TG) and VLDL-C of serum (p<0.05). Levels of thiobarbituric acid reactive substance (TBARS) in the liver tended to decrease with the increasing of the P/S ratio in the diet (p<0.001), but antioxidant enzyme activity in the liver was not significantly different. In addition, antioxidant supplementation significantly lowered TBARS level in the liver (p<0.05), but had no effect on antioxidant enzyme activity except for glutathione reductase (p<0.05). In conclusion, it is necessary to consider the properties of dietary fatty acids and antioxidants supplementation for the prevention of cardiovascular diseases.