In the present study, the concentration of NG,NG-dimethylarginine (DMA), an endogenous inhibitor of nitric oxide synthesis, was measured with high-performance liquid chromatography, and the effect of hypercholesterolemia on DMA level was investigated in the high fat, high cholesterol fed rabbit. After 6 weeks on a high fat, high cholesterol diet, serum total cholesterol, triglycerides, and lipid peroxides were increased, and atherosclerosis was shown by means of morphological examination. In these rabbits with atherosclerosis, serum level of DMA was significantly raised, while serum creatine level remained normal. This study suggests that chronic hypercholesteremia may stimulate DMA production through elevation of lipid peroxides. The present results implicate endogenous DMA as a contributor to the development of atherosclerosis.