Pumpkin-seed oil (PSO), a natural supplement rich with antioxidants and polyunsaturated fatty acids (PUFAs), was given in combination with simvastatin, as antihypercholesterolemic drug, to high cholesterol-fed rabbits, for three weeks. In comparison with normal rabbits, a significant increase of the aortic contractile response to norepinephrine was observed which could be attributed to endothelium dysfunction. In addition, serum levels of total lipids, triacylglycerols, total cholesterol and low density lipoprotein-cholesterol (LDL-C) were increased while phospholipids and high density lipoprotein-cholesterol (HDL-C) were decreased in hypercholesterolemic rabbits. These changes could be related to the predominance of LDL and oxidized-LDL particles caused by high levels of reactive oxygen species during hypercholesterolemia (HC). Treatment with simvastatin modulated most of the altered parameters affected during HC that might be, in part, due to inhibition of cholesterol biosynthesis. While concomitant administration of simvastatin and PSO succeeded to cause marked reduction of the aortic contractile response to norepinephrine and to normalize the most adverse effects observed during HC. These effects were explained by the potentiating effects of simvastatin with antioxidants and essential fatty acids in PSO. On the contrary, serum activities of aminotransferases and creatine phosphokinase were increased with simvastatin treatment but not with the combination therapy in hypercholesterolemic rabbits.