17 beta-estradiol regulation of human endothelial cell basal nitric oxide release, independent of cytosolic Ca2+ mobilization.
17 beta-estradiol regulation of human endothelial cell basal nitric oxide release, independent of cytosolic Ca2+ mobilization.
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DOI：
10.1161/01.res.81.5.885
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发表时间：
1997-11
影响因子：
20.1
通讯作者：
T. Caulin–Glaser;G. Garcı́a-Cardeña;P. Sarrel;W. Sessa;J. Bender
中科院分区：
文献类型：
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作者：
T. Caulin–Glaser;G. Garcı́a-Cardeña;P. Sarrel;W. Sessa;J. Bender
雌二醇可延缓动脉粥样硬化的发展。动物模型表明，NO可能是这种心血管保护的关键效应分子。在这项研究中，女性人脐静脉内皮细胞（HUVECs）增殖的酚红无性腺激素的培养基和预处理的17 β-雌二醇（E2）。减少NO2-和NO3-（NOx）的浓度，通过化学发光测定，表现出快速增加基础HUVEC NO释放响应生理浓度的E2。雌激素受体（ER）拮抗剂ICI 164，384抑制了NO释放的增加，证明了这种反应的ER介导的成分。由于内皮型一氧化氮合酶（eNOS）的活性在很大程度上是由胞质Ca 2+调节，相对[Ca 2 +]i响应于E2测定在荧光测定。E2对HUVEC Ca ~（2+）通量无促进作用。此外，eNOS活性在E2预处理的内皮全细胞裂解物是不依赖于额外的Ca 2+。尽管涉及ER，但这是一种非基因组效应E2，如转录抑制细胞中维持的反应和NO生物测定中cGMP快速（10分钟）形成所证明的。我们证明，第一次，独立的胞质Ca 2+动员，有增强eNOS活性，从而增加在HUVEC基础NO释放响应短期雌二醇暴露。雌激素的心血管保护作用的影响进行了讨论。
Estradiol retards the development of atherosclerosis. Animal models have suggested that NO may be a critical effector molecule in this cardiovascular protection. In this study, female human umbilical vein endothelial cells (HUVECs) were propagated in phenol red-free gonadal hormone-free medium and pretreated with 17 beta-estradiol (E2). Reduced NO2- and NO3- (NOx) concentration, determined by chemiluminescence, demonstrated a rapid increase in basal HUVEC NO release in response to physiological concentrations of E2. The estrogen receptor (ER) antagonist ICI 164,384 inhibited the augmented NO release, demonstrating an ER-mediated component of this response. Because endothelial NO synthase (eNOS) activity is largely regulated by cytosolic Ca2+, relative [Ca2+]i in response to E2 was determined in a fluorometric assay. E2 did not promote HUVEC Ca2+ fluxes. Furthermore, eNOS activity in E2-pretreated endothelial whole-cell lysates was not dependent on additional Ca2+. Despite involving the ER, this is a nongenomic effect E2, as demonstrated by maintained responses in transcriptionally inhibited cells and by the rapidly (10 minutes) of cGMP formation in an NO bioassay. We demonstrate, for the first time, that independent of cytosolic Ca2+ mobilization, there is augmentation of eNOS activity with a resultant increase in HUVEC basal NO release in response to short-term estradiol exposure. Implications for the cardiovascular protective role of estrogen are discussed.