Catecholamines Abrogate the Anti-Mitogenic Effects of 2-Hydroxy Metabolite of Estradiol on Vascular Smooth Muscle Cells by Inhibiting Catechol-O-Methyltransferase (COMT) Activity and 2-Methoxyestradiol Formation
Methylation of 2-hydroxyestradiol(2OHE; endogenous estradiol metabolite) to 2-methoxyestradiol (2MeOE; angiogenesis inhibitor)by COMT plays a key role in mediating the anti-mitogenic effects of 2OHE on vascular smooth muscle cell (SMC)growth. Catecholamines such as norepinephrine (NE) are also substrates for COMT and increased levels of NE are associated with vasoocclusive disorders. We hypothesize that increased endogenous synthesis/levels of NE under pathophysiological conditions may abrogate the vasoprotective effects of 2OHE by competing for COMT and inhibiting 2MeOE formation. To test this hypothesis we investigated the anti-mitogenic effects of .001-10μM 2OHE on 2.5% FCS-induced SMC growth (cell number, DNA synthesis [thymidine incorporation], collagen synthesis [proline incorporatio])in rat and human aortic SMCs in the presence and absence of NE (0.1-40μM). NE concentration-dependently abrogated the inhibitory effects of 2OHE on SMC growth and in the presence of 10μM NE the inhibitory curve of 2OHE on SMC growth was shifted to the right(P<.05). In the presence of 10μM NE, the inhibitory effect of 1μM 2OHE on DNA synthesis was reduced from 70±3% to 24±2% (P<.05), and this effect of NE was mimicked by isoproterenol (ISO) and epinephrine (EPI). Additionally, NE (0.5-2.5mM) inhibited the metabolism of 10μM 2OHE to 2MeOE in a concentration-dependent manner and the effects of NE were mimicked by ISO, EPI, metanephrine, normetanephrine and 3,4-dihydroxymandelic acid. At 0.5 mM ISO, NE and EPI inhibited 2MeoE formation by 70±4%,20±2% and 40±2%, respectively. Our findings suggest that increases in local synthesis of catecholamines within the vasculature may abrogate the anti-vasoocclusive effects of estradiol and 2OHE by blocking 2MeOE formation. In conclusion, the interaction between catecholamines and 2OHE may play a key role in the biology of vascular SMC growth.