The impact of the source of n-3 fatty acids on cardiovascular disease has not been fully investigated. This study was carried out to investigate cardiovascular benefits of diets with a low ratio (2:1) of n-6:n-3 fatty acids from different sources, either fish or flaxseed oil, in C57BL/6 mice. Twenty-one mice were divided into 3 groups (n=7) and fed an atherogenic diet supplemented with either a fish or flaxseed oil-based ‘designer oil’ with low n-6:n-3 fatty acid ratio (treated groups) or safflower oil-based formulation with a high ratio (control group) for 16 weeks. Plasma cholesterol levels declined significantly in both treated groups, by greater than 30%, compared to those in control. The ratio of n-6:n-3 fatty acids in liver was significantly lower in fish and flax groups as compared to control. Our data suggest that lowering dietary ratio of n-6:n-3 fatty acids may significantly reduce cardiovascular risks regardless of the source of n-3 fatty acids.