Aim To observe the effect of Shenshao oral lotion on serum lipids and vasoactive substances in atherosclerotic rats. Methods Atherosclerotic rats model were established by high fat feed and intraperitoneal injection of vitamin D3. 60 male SD rats were divided into six groups randomly: normal feed group, atherosclerotic model group, low dose of Shenshao oral lotion group, middle dose of Shenshao oral lotion group and high dose of Shenshao oral lotion group, simvastatin control group. Pathomorphology change of aorta of the rats was observed through light microscope. Serum lipids and vasoactive substances level of rats in each groups were examined after 12 weeks feed and Shenshao oral lotion were given for 4 weeks. Results The pathological change of aorta of the rats in high dose of Shenshao oral lotion group was alleviated compared with that in atherosclerotic model group. Compared with atherosclerotic model groups, the serum level of total cholesterol, triglyceride, low density lipoprotein cholesterol, endothelin, angiotensinⅡ, thromboxane B2 were lower and the serum level of high density lipoprotein cholesterol, nitric oxide synthase were higher in high dose Shenshao oral lotion group and simvastatin control group（P<0.05）. Conclusions Shenshao oral lotion could modulate blood lipids and vasoactive substances concentration in experimental rats, which may be one of the mechanism of Shenshao oral lotion to prevent atherosclerosis.