Estradiol-17β and Its Cytochrome P450- and Catechol- O -Methyltransferase–Derived Metabolites Stimulate Proliferation in Uterine Artery Endothelial Cells

Abstract

Abstract-Estradiol-17␤ (E 2 ␤) and its metabolites, which are sequentially synthesized by cytochrome P450s and catechol-O-methyltransferase to form 2 and 4-hydroxyestradiol (OHE 2 ) and 2-and 4-methoxestradiol (ME 2 ), are elevated during pregnancy. We investigated whether cytochrome P450s and catechol-O-methyltransferase are expressed in uterine artery endothelial cells (UAECs) and whether E 2 ␤ and its metabolites modulate cell proliferation via ER-␣ and/or ER-␤ and play roles in physiological uterine angiogenesis during pregnancy. Cultured ovine UAECs from pregnant and nonpregnant ewes were treated with 0.1 to 100.0 nmol/L of E 2 ␤, 2-OHE 2 , 4-OHE 2 , 2-ME 2 , and 4-ME 2 . ER-␣ or ER-␤ specificity was tested using ICI 182 780, ER-␣-specific 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinyleth oxy)phenol]-1H-pyrazole dihydrochloride, ER-␤-specific 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo [1,5-a]pyrim idin-3-yl]phenol antagonists and their respective agonists ER-␣-specific 4,4Ј,4Љ-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol and ER-␤-specific 2,3-bis(4-Hydroxyphenyl)-propionitrile. Angiogenesis was evaluated using 5-bromodeoxyuridine proliferation assay. Using confocal microscopy and Western analyses to determine enzyme location and levels, we observed CYP1A1, CYP1A2, CYP1B1, CYP3A4, and catechol-O-methyltransferase expression in UAECs; however, expressions were similar between nonpregnant UAECs and pregnant UAECs. E 2 ␤, 2-OHE 2 , 4-OHE 2 , and 4-ME 2 treatments concentration-dependently stimulated proliferation in pregnant UAECs but not in nonpregnant UAECs; 2-ME 2 did not stimulate proliferation in either cell type. Proliferative responses of pregnant UAECs to E 2 ␤ were solely mediated by ER-␤, whereas responses to E 2 ␤ metabolites were neither ER-␣ nor ER-␤ mediated. We demonstrate an important vascular role for E 2 ␤, its cytochrome P450-and catechol-O-methyltransferasederived metabolites, and ER-␤ in uterine angiogenesis regulation during pregnancy that may be dysfunctional in preeclampsia and other cardiovascular disorders. (Hypertension. 2010;55:1005-1011.)Key Words: angiogenesis Ⅲ hypertension Ⅲ pregnancy Ⅲ endothelium Ⅲ estradiol metabolites Ⅲ CYP450s P regnancy is associated with dramatic uterine blood flow rises resulting from vascular adaptations, including vasodilatation and angiogenesis. 1 These adaptations are critical in pregnancy because their dysfunctions are implicated in pathological pregnancies, such as preeclampsia, which complicates 6% to 8% of all pregnancies in the United States and accounts for 50 000 maternal deaths per year worldwide. [2][3][4] Regulation of vascular adaptations during pregnancy is mediated partly by estrogens, which are elevated during gestation. 5 Estradiol-17␤ (E 2 ␤) infusion in sheep markedly reduces uterine and systemic vascular resistance causing rises in uterine and systemic blood flows. 6 Uterine arterial administration of the nonselective estrogen receptor (ER) antagonist ICI 182 780 (ICI) in pregnant sheep lowers uterine blood flow, demonstra...

Uterine artery leptin receptors during the ovarian cycle and pregnancy regulate angiogenesis in ovine uterine artery endothelial cells†

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The Pharmacology of the Cytochrome P450 Epoxygenase/Soluble Epoxide Hydrolase Axis in the Vasculature and Cardiovascular Disease

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