Abstract-Catechol-O-methyltransferase (COMT)-mediated methylation of 2-hydroxyestradiol (endogenous estradiol metabolite) to 2-methoxyestradiol (angiogenesis inhibitor) may be responsible for the antimitogenic effects of 2-hydroxyestradiol on vascular smooth muscle cells (VSMCs). Catecholamines are also substrates for COMT, and increased levels of catecholamines are associated with vasoocclusive disorders. We hypothesize that catecholamines may abrogate the vasoprotective effects of 2-hydroxyestradiol by competing for COMT and inhibiting 2-methoxyestradiol formation. To test this hypothesis, we investigated the antimitogenic effects of 0.001 to 0.1 mol/L of 2-hydroxyestradiol on human aortic VSMC proliferation (cell number and DNA synthesis), collagen synthesis, and migration in the presence and absence of catecholamines. Norepinephrine, epinephrine, and isoproterenol concentrationdependently abrogated the inhibitory effects of 2-hydroxyestradiol on cell number, DNA synthesis, collagen synthesis, and cell migration. These modulatory/attenuating effects of catecholamines were not abrogated in the presence of the ␣-and -adrenergic receptor antagonists, phentolamine mesylate and propranolol, respectively. In contrast to 2-hydroxyestradiol, the antimitogenic effects of 2-methoxyestradiol (0.1 mol/L) were not attenuated by isoproterenol (1 mol/L) or quercetin (competitive inhibitor of COMT, 10 mol/L). Norepinephrine, epinephrine, and isoproterenol concentration-dependently (10 to 500 mol/L) inhibited the metabolism of 2-hydroxyestradiol (0.25 to 2 mol/L) to 2-methoxyestradiol, and the potency of the catecholamines to reverse 2-hydroxyestradiol-induced inhibition of VSMC proliferation, collagen synthesis, and migration was correlated with their ability to inhibit 2-methoxyestradiol formation. Key Words: estradiol Ⅲ catechol estrogens Ⅲ methoxyestradiol Ⅲ menopause Ⅲ vascular remodeling E stradiol induces anti-vaso-occlusive effects in some, but not all, women. 1 Because estradiol prevents injuryinduced neointimal formation in mice lacking functional estrogen receptors (ERs), 2,3 the anti-vaso-occlusive effects of estradiol may in part be mediated by ER-independent mechanisms. In this regard, we have recently shown that the antimitogenic effects of estradiol on vascular smooth muscle cell (VSMC) growth are mediated via its metabolites with little or no affinity for ERs. 4 We hypothesize that 2-hydroxyestradiol mediates, in part, the cardiovascular protective effects of estradiol. In support of this hypothesis, our previous findings show that 2-hydroxyestradiol, a major endogenous metabolite of estradiol with little affinity for ERs, is more potent than estradiol in inhibiting VSMC growth. 4,5 We also hypothesize that the growth-inhibitory effects of 2-hydroxyestradiol are mediated mostly via 2-methoxyestradiol. Indeed, 2-methoxyestradiol, the methylated product of 2-hydroxyestradiol with no binding affinity for the ERs, is even more potent than is 2-hydroxyestradiol in inhibiting cell growth. 4,6 Moreover, our prior s...