This study examined the effects of fish protein in combination with fish oil on rat lipid metabolism. Male Wistar rats were divided into four groups and fed an AIN93G-based diet with casein (20%) + soybean oil (7%), casein (10%) + fish protein (10%) + soybean oil (7%), casein (20%) + soybean oil (5%) + fish oil (2%), and casein (10%) + fish protein (10%) + soybean oil (5%) + fish oil (2%) for 4 weeks. The dietary combination of fish protein and fish oil decreased the contents of serum triacylglycerol, serum cholesterol, liver triacylglycerol and liver cholesterol in addition to altering liver lipid fatty acid composition. These effects are partly due to the increase in fecal cholesterol, bile acid excretion, and increased enzyme activities of fatty acid β-oxidation in the liver. These data suggest that combined intake of fish protein and fish oil lead to both hypocholesterolemic and hypotriglyceridemic in serum and the liver, while sole intake of fish protein or fish oil decrease only cholesterol and triglyceride levels, respectively. These results suggest that combined intake of fish protein and fish oil may play beneficial roles in the prevention of lifestyle-related diseases as compared with sole fish protein intake.