The mechanisms of the protective effect of 17B-estradiol were investigated in the middle cerebral artery (MCA) and aorta isolated from cholesterol-fed rabbits. Three groups were assigned: control group (standard chow), cholesterol group (standard chow+1% cholesterol) and estradiol group (1% cholesterol+17B-estradiol). The MCA and the aorta were isolated, precontracted respectively with high K(+) solution or with phenylephrine and exposed to cumulative acetylcholine concentrations. In the control group, acetylcholine induced a concentration-dependent relaxation in the aorta and the MCA. Cholesterol diet for eight months reduced significantly the maximal response to acetylcholine by about 50% in the aorta and by about 30% in the MCA. The chronic treatment with 17B-estradiol restored this impaired relaxations to acetylcholine. Incubation of arteries from estradiol group with N(omega)-nitro L-arginine methyl-ester (L-NAME), a potent inhibitor of constitutive nitric oxide synthase, entirely abolished the relaxation to acetylcholine while aminoguanidine, a potent inhibitor of inducible nitric oxide synthase, did not affect this relaxation. These observations suggest that the protective effect of 17B-estradiol against hypercholesterolemia is mediated via a release of endothelial nitric oxide.