Estrogen Replacement Therapy in Ovariectomized Nonpregnant Ewes Stimulates Uterine Artery Hydrogen Sulfide Biosynthesis by Selectively Up-Regulating Cystathionine β-Synthase Expression
Estrogen Replacement Therapy in Ovariectomized Nonpregnant Ewes Stimulates Uterine Artery Hydrogen Sulfide Biosynthesis by Selectively Up-Regulating Cystathionine β-Synthase Expression
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DOI：
10.1210/en.2015-1086
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发表时间：
2015-06-01
期刊：
影响因子：
4.8
通讯作者：
Chen, Dong-bao
中科院分区：
文献类型：
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作者：
Lechuga, Thomas J.;Zhang, Hong-hai;Chen, Dong-bao
雌激素显著扩张许多血管床，子宫内反应最大。内源性硫化氢（H2S）是一种有效的血管扩张剂和促血管生成的第二信使，由l -半胱氨酸通过胱硫氨酸-合成酶（CBS）和胱硫氨酸-裂解酶（CSE）合成。我们假设雌激素替代疗法（ERT）选择性地刺激子宫动脉（UA）和其他全身动脉中H2S的生物合成。从接受载体或雌二醇-17 β替代疗法（ERT）的切除卵巢的未怀孕母羊（n = 5/组）中获得完整的和内皮脱落的UA、肠系膜动脉（MA）和颈动脉（CA）。提取总RNA和总蛋白，测定CBS和CSE，亚甲基蓝法测定H2S产量。免疫荧光显微镜下，用石蜡包埋的UA环定位CBS和CSE蛋白。ERT显著刺激CBS mRNA和蛋白，而未改变完整和剥落UA的CSE mRNA和蛋白。定量免疫荧光显微镜分析显示cbsa和CSE蛋白在内皮平滑肌中定位，并证实ERT刺激CBS而不是UA内皮和平滑肌中CSE蛋白的表达。ERT也刺激了去卵巢母羊的CBS，但没有刺激CSE、mRNA和蛋白的表达，但没有刺激CA的表达。同时，ERT刺激了UA和ma，而不是CA H2S的产生。一种特定的CBS可以完全阻断ert刺激下的UA H2S生成，而CSE抑制剂则不能。因此，ERT通过特异性上调CBS表达，选择性地刺激UA和MA，而不是CA H2S的生物合成，暗示H2S在雌激素诱导的血管舒张和绝经后妇女健康中的作用。
Estrogens dramatically dilate numerous vascular beds with the greatest response in the uterus. Endogenous hydrogen sulfide (H2S) is a potent vasodilator and proangiogenic second messenger, which is synthesized from L-cysteine by cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CSE). We hypothesized that estrogen replacement therapy (ERT) selectively stimulates H2S biosynthesis in uterine artery (UA) and other systemic arteries. Intact and endothelium-denuded UA, mesenteric artery (MA), and carotid artery (CA) were obtained from ovariectomized nonpregnant ewes (n = 5/group) receiving vehicle or estradiol-17 beta replacement therapy (ERT). Total RNA and protein were extracted for measuring CBS and CSE, and H2S production was determined by the methylene blue assay. Paraffin-embedded UA rings were used to localize CBS and CSE proteins by immunofluorescence microscopy. ERT significantly stimulated CBS mRNA and protein without altering CSE mRNA or protein in intact and denuded UA. Quantitative immunofluorescence microscopic analysesshowedCBSandCSE protein localization in endotheliumandsmoothmuscleand confirmed that ERT stimulated CBS but not CSE protein expression in UA endothelium and smooth muscle. ERT also stimulated CBS, but not CSE, mRNA and protein expression in intact and denuded MAbut not CA in ovariectomized ewes. Concomitantly, ERT stimulated UA andMAbut not CA H2S production. ERT-stimulated UA H2S production was completely blocked by a specific CBS but not CSE inhibitor. Thus, ERT selectively stimulates UA and MA but not CA H2S biosynthesis by specifically up-regulating CBS expression, implicating a role of H2S in estrogen-induced vasodilation and postmenopausal women's health.