Li Wan-Li, Xiang Wei, Ping Ye
Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China.
Mol Med Rep. 2015 Aug;12(2):2425-30. doi: 10.3892/mmr.2015.3674. Epub 2015 Apr 24.
Several studies have recently demonstrated that G protein-coupled estrogen receptor (GPER) 30 directly binds to estrogen and mediates its action. The aim of the present study was to investigate the effects of GPER on cardiocyte apoptosis following ischemia/reperfusion injury (MIRI) in H9C2 myocardial cells. H9C2 cells were treated with a specific GPER agonist (G1), 17β-estradiol (E2) or the vehicle. The cells were subjected to 20 min of myocardial ischemia followed by 120 min of reperfusion. They were then randomly assigned to three experimental groups: Control, G1, E2. B-cell lymphoma 2 (Bcl-2) and Bcl-2 associated X (Bax) levels were measured, Hoechst 33258 staining was performed to assess apoptosis, and superoxide dismutase (SOD), tumor necrosis factor (TNF)-α and adenosine triphosphatase (ATPase) levels were determined. To test the specificity of G1, GPER-knockout cells were treated with G1 and analyzed as stated above. Compared with the vehicle-treated groups, G1 and E2-treated groups exhibited elevated Bcl-2 levels, decreased Bax levels and cell apoptosis, significantly increased SOD and ATP levels and decreased TNF-α levels following ischemia-reperfusion. However, G1 had no evident effects on the GPER-knockout cells. In conclusion, the present study suggested that GPER activation provided a cardioprotective effect following ischemia-reperfusion by inhibiting cardiocyte apoptosis.
最近的几项研究表明，G蛋白偶联雌激素受体（GPER）30可直接与雌激素结合并介导其作用。本研究的目的是探讨GPER对H9C2心肌细胞缺血/再灌注损伤（MIRI）后心肌细胞凋亡的影响。用特异性GPER激动剂（G1）、17β-雌二醇（E2）或赋形剂处理H9C2细胞。使细胞经历20分钟的心肌缺血，随后进行120分钟的再灌注。然后将它们随机分为三个实验组：对照组、G1组、E2组。检测B细胞淋巴瘤2（Bcl-2）和Bcl-2相关X蛋白（Bax）水平，进行Hoechst 33258染色以评估细胞凋亡，并测定超氧化物歧化酶（SOD）、肿瘤坏死因子（TNF）-α和三磷酸腺苷酶（ATPase）水平。为了测试G1的特异性，用G1处理GPER基因敲除细胞，并按上述方法进行分析。与赋形剂处理组相比，G1组和E2组在缺血再灌注后Bcl-2水平升高，Bax水平降低，细胞凋亡减少，SOD和ATP水平显著升高，TNF-α水平降低。然而，G1对GPER基因敲除细胞没有明显影响。总之，本研究表明，激活GPER可通过抑制心肌细胞凋亡对缺血再灌注后的心脏起到保护作用。
