Aim To observe the effect of probucol on endothelium-dependent vasodilator activity in high cholesterol plus high fat diet (HCHF) rabbits. Methods 26 New Zealand white rabbits (1.5-2.5 kg) were randomly divided into three groups, that is, control group ( n = 6, normal diet); hyperlipidemic group ( n = 10, HCHF-diet); probucol group ( n = 10, HCHF-diet plus probucol 200 mg/kg everyday). There blood was drawn from the central ear artery to analyze plasma total cholesterol (TC) at the pre-experiment and in the 12th week of post experiment. Then all rabbits were killed in the 12 th week and thoracic aortas were isolated to analyze vascular endothelial function. Results Probucol could remarkably lower plasma TC lever,compared to hyperlipidemic group, TC level decreased 39.1 % in probucol group ( P< 0.01). The HCHF diet significantly attenuated endothelium-dependent relaxation (EDR) of rabbit aortic rings to acetylcholine (Ach), but probucol could significantly protected against HCHF diet-injured EDR of rabbit aortic rings. The HCHF diet and probucol did not significantly influence the endothelium-independent relaxation of the rabbit artery elicited by sodium nitroprusside (SNP). Conclusion Probucol can protect against injuries of vascular endothelial function induced by hypercholesterolemia through lowering plasma TC and anti-oxidant.