ERα/ERβ-directed CBS transcription mediates E2β-stimulated hUAEC H2S production
ERα/ERβ-directed CBS transcription mediates E2β-stimulated hUAEC H2S production
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DOI：
10.1530/jme-22-0175
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发表时间：
2023-02-01
影响因子：
3.5
通讯作者：
Chen, Dong-bao
中科院分区：
文献类型：
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作者：
Bai, Jin;Lechuga, Thomas J.;Chen, Dong-bao
升高的内源性雌激素通过选择性上调H2S合成酶胱硫醚β-合酶（CBS）的表达来刺激人子宫动脉内皮细胞（hUAEC）产生硫化氢（H2S），但其潜在机制尚不清楚。我们假设 CBS 转录介导雌激素刺激的妊娠依赖性 hUAEC H2S 产生。雌二醇 17 β (E-2 β) 刺激离体妊娠人子宫动脉中的 CBS，但不刺激胱硫醚 γ 裂解酶 (CSE) 的表达，雌激素受体 (ER) 拮抗剂 ICI 182,780 可减弱这种作用。 E-2 beta 刺激非妊娠和妊娠女性原代 hUAEC 中 CBS mRNA/蛋白和 H2S 的产生，但在妊娠状态下反应更大；全部被 ICI 182,780 屏蔽。人CBS启动子含有多种雌激素反应元件(ERE)，包括1个优先结合ER α的ERE(α ERE)和3个优先结合ER β的ERE(β ERE)，以及1个全ERE(α/β ERE)和1个半ERE(1/2 α/β ERE)同时结合ER α和ER β。使用由具有一系列 5 & PRIME;-缺失的人类 CBS 启动子驱动的报告基因进行荧光素酶测定，确定结合 ER α 和 ER β 的 α/β ERE（α/β ERE 和 1/2 α/β ERE）对于基线和 E-2 β 刺激的 CBS 启动子激活非常重要。 E-2β通过募集ERα到α/βERE和βERE，以及将ERβ募集到βERE、α/βERE和αERE，刺激ERα/ERβ异二聚化。单独的ERα或ERβ激动剂反式激活CBS启动子，刺激CBS mRNA/蛋白和H2S产生至与E-2β刺激的水平相当的水平，而单独的ERα或ERβ拮抗剂消除E-2β刺激的反应。 E-2 beta 不会改变 hUAEC 中的人 CSE 启动子活性和 CSE mRNA/蛋白。总而言之，雌激素刺激的妊娠依赖性 hUAEC H2S 产生是通过 ER α/ER β 指导的基因转录选择性上调 CBS 表达而发生的。
Elevated endogenous estrogens stimulate human uterine artery endothelial cell (hUAEC) hydrogen sulfide (H2S) production by selectively upregulating the expression of H2S synthesizing enzyme cystathionine beta-synthase (CBS), but the underlying mechanisms are underdetermined. We hypothesized that CBS transcription mediates estrogen-stimulated pregnancy-dependent hUAEC H2S production. Estradiol-17 beta (E-2 beta) stimulated CBS but not cystathionine gamma-lyase (CSE) expression in pregnant human uterine artery ex vivo, which was attenuated by the estrogen receptor (ER) antagonist ICI 182,780. E-2 beta stimulated CBS mRNA/protein and H2S production in primary hUAEC from nonpregnant and pregnant women, but with greater responses in pregnant state; all were blocked by ICI 182,780. Human CBS promoter contains multiple estrogen-responsive elements (EREs), including one ERE preferentially binding ER alpha (alpha ERE) and three EREs preferentially binding ER beta (beta ERE), and one full ERE (alpha/beta ERE) and one half ERE (1/2 alpha/beta ERE) binding both ER alpha and ER beta. Luciferase assays using reporter genes driven by human CBS promoter with a series of 5 & PRIME;-deletions identified the alpha/beta EREs binding both ER alpha and ER beta (alpha/beta ERE and 1/2 alpha/beta ERE) to be important for baseline and E-2 beta-stimulated CBS promoter activation. E-2 beta stimulated ER alpha/ER beta heterodimerization by recruiting ER alpha to alpha/beta EREs and beta ERE, and ER beta to beta ERE, alpha/beta EREs, and alpha ERE. ER alpha or ER beta agonist alone trans-activated CBS promoter, stimulated CBS mRNA/protein and H2S production to levels comparable to that of E-2 beta-stimulated, while ER alpha or ER beta antagonist alone abrogated E-2 beta-stimulated responses. E-2 beta did not change human CSE promoter activity and CSE mRNA/protein in hUAEC. Altogether, estrogen-stimulated pregnancy-dependent hUAEC H2S production occurs by selectively upregulating CBS expression via ER alpha/ER beta-directed gene transcription.