In the present study, we examined the effects of fish oil supplementation in 3 months old male Wistar rats on changes in plasma and liver lipid metabolism and oxidative stress parameters. Twenty Wistar rats were randomly divided into two groups of ten animals: control group and intervention group, treated for 6 weeks with fish oil capsules containing 45 mg eicosapentanoic acid and 30 mg docosahexanoic acid. After intervention, biochemical parameters in plasma [triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL) and total cholesterol, urea, creatinine and uric acid], fatty acid (FAs) profile of liver phospholipids and parameters of oxidative stress in liver [activity of catalase, superoxide dismutase and paraoxonase (PON1), concentration of nitrites, lipid peroxidation (LPO), free thiol (SH) groups and lactate dehydrogenase (LDH) izoenzymes were determined. Treatment with fish oil improved FAs profile of liver phospholipids, increasing n-3 FAs and decreasing n-6/n-3 ratio. Significant decrease in plasma TG and LDL concentration, and increase in the level of HDL and uric acid were found in intervention group at the end of the study. Catalase activity, LPO, and nitrites concentration in liver were significantly decreased, after the supplementation, together with elevated PON1 activity. Applied treatment significantly improved plasma lipid profile, liver FAs composition and parameters of oxidative stress in male Wistar rats.