Estrogen receptor-beta activation results in S-nitrosylation of proteins involved in cardioprotection.
Estrogen receptor-beta activation results in S-nitrosylation of proteins involved in cardioprotection.
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DOI：
10.1161/circulationaha.109.868729
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发表时间：
2009-07-21
期刊：
影响因子：
37.8
通讯作者：
Sun J
中科院分区：
文献类型：
--
作者：
Lin J;Steenbergen C;Murphy E;Sun J
已有研究表明，雌激素受体-β(ER-β)的激活对缺血再灌注损伤具有重要的心脏保护作用。然而，这种保护的机制尚不清楚。我们假设雌激素通过激活ER-β来保护心脏，这导致了关键的心脏保护蛋白的S亚硝酸化。我们用ER-β选择性激动剂2，2-二(4-羟基苯基)-丁腈、17-β-雌二醇(E_2)或用阿尔茨海默特微泵治疗去卵巢的C57BL/6J小鼠两周。离体心经Langendorff灌流后进行缺血再灌流。与赋形剂处理的心脏相比，DPN和E2处理的心脏在缺血后的功能恢复和缩小梗塞面积方面有明显的改善。为了检测DPN的特异性，我们用DPN处理ER-β基因敲除(βERKO)小鼠。然而，在βERKO小鼠中未发现该药的心脏保护作用，提示该药的心脏保护作用是通过激活ER-β来实现的。利用DyLight-Maleimide荧光剂和一种改进的生物素开关方法，我们使用了二维DyLight荧光差异凝胶电泳法(2D DyLight Dige)蛋白质组学方法来定量蛋白质SNO的差异。DPN和E2处理的心脏显示出许多蛋白质的SNO增加。有趣的是，这些蛋白质中的许多也被证明在预适应的心脏中增加了SNO。此外，一氧化氮合酶(NO)抑制剂可消除DPN诱导的心脏保护作用和增加的SNO。结论：雌激素可通过激活ER-β和NO/β信号转导途径来保护心肌，从而增加心肌细胞的S亚硝化水平。
It has been shown that the activation of estrogen receptor-β (ER-β) plays an important cardioprotective role against ischemia-reperfusion (I/R) injury. However, the mechanism for this protection is not clear. We hypothesize that estrogen protects by ER-β activation, which leads to S-nitrosylation (SNO) of key cardioprotective proteins. We treated ovariectomized C57BL/6J mice with an ER-β selective agonist, 2,2-bis(4-hydroxyphenyl)-proprionitrile (DPN), 17β-estradiol (E2) or vehicle using Alzet minipumps for two weeks. Isolated hearts were Langendorff perfused and subjected to ischemia and reperfusion. Compared with vehicle-treated hearts, DPN and E2-treated hearts had significantly better post-ischemic functional recovery and decreased infarct size. To test the specificity of DPN, we treated ER-β knockout (βERKO) mice with DPN. However, no cardioprotective effect of DPN was found in βERKO mice, indicating the DPN-induced cardioprotection occurs through the activation of ER-β. Using DyLight-maleimide fluors and a modified biotin switch method, we employed a two dimensional DyLight fluorescence difference gel electrophoresis (2D DyLight DIGE) proteomic method to quantify differences in SNO of proteins. DPN and E2-treated hearts showed an increase in SNO of a number of proteins. Interestingly, many of these proteins had also been shown to have increased SNO in preconditioned hearts. In addition, the DPN-induced cardioprotection and increased SNO were abolished by treatment with a nitric oxide (NO) synthase inhibitor. The activation of ER-β by DPN treatment leads to increased protein S-nitrosylation and cardioprotection against I/R injury, suggesting that chronic estrogen exposure protects hearts largely via activation of ER-β and NO/SNO signaling.