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首页> 外文期刊>Cell biology international. >Calcium-sensing receptors induce apoptosis in cultured neonatal rat ventricular cardiomyocytes during simulated ischemia/reperfusion.
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Calcium-sensing receptors induce apoptosis in cultured neonatal rat ventricular cardiomyocytes during simulated ischemia/reperfusion.

机译:在模拟的缺血/再灌注过程中,钙敏感受体在培养的新生大鼠心室心肌细胞中诱导凋亡。

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Calcium-sensing receptors (CaSRs) are G-protein coupled receptors which regulate systemic calcium homeostasis and also participate in cell proliferation, differentiation and apoptosis. We have previously shown that CaSR can induce apoptosis in isolated rat adult hearts and in normal rat neonatal cardiomyocytes. However, no knowledge exists concerning the role of CaSR in apoptosis induced by ischemia and reperfusion in neonatal cardiac myocytes. Therefore, in the present study, we incubated primary neonatal rat ventricular cardiomyocytes in ischemia-mimetic solution for 2h, then re-incubated them in a normal culture medium for 24h to establish a model of simulated ischemia/reperfusion (I/R). We assayed the apoptotic ratio of the cardiomyocytes by flow cytometry; observed morphological alterations by transmission electron microscope; analyzed the expression of caspase-3, Bcl-2, CaSR, extracellular signal-regulated protein kinase (ERK), and Fas/Fas ligand (FasL) by Western blotting; and measured the concentration of intracellular calcium by Laser Confocal Scanning Microscopy. The results showed that simulated I/R increased the expression of CaSR and cardiomyocyte apoptosis. GdCl(3), a specific activator of CaSR, further enhanced CaSR expression, along with increases in intracellular calcium and apoptosis in cardiomyocytes during I/R. Activation of CaSR down-regulated Bcl-2 expression, up-regulated caspase-3 and Fas/FasL expression and stimulated ERK1/2 phosphorylation. In summary, CaSR is involved in I/R injury and apoptosis of neonatal rat ventricular cardiomyocytes by inhibiting Bcl-2, inducing calcium overload and activating the Fas/FasL death receptor pathway.
机译:钙敏感受体(CaSR)是G蛋白偶联受体,可调节全身钙稳态,并参与细胞增殖,分化和凋亡。先前我们已经表明,CaSR可以诱导离体大鼠成年心脏和正常大鼠新生心肌细胞凋亡。然而,关于CaSR在新生心肌细胞的局部缺血和再灌注诱导的凋亡中的作用尚无知。因此,在本研究中,我们将原代新生大鼠心室心肌细胞在模拟缺血溶液中孵育2h,然后在正常培养基中将其重新孵育24h,以建立模拟缺血/再灌注(I / R)模型。我们通过流式细胞术测定了心肌细胞的凋亡率。通过透射电子显微镜观察形态变化;通过蛋白质印迹分析了caspase-3,Bcl-2,CaSR,细胞外信号调节蛋白激酶(ERK)和Fas / Fas配体(FasL)的表达;通过激光共聚焦扫描显微镜测定细胞内钙的浓度。结果表明,模拟的I / R增加了CaSR的表达和心肌细胞凋亡。 GdCl(3),CaSR的特定激活剂,进一步增强了CaSR的表达,以及I / R期间心肌细胞内钙的增加和心肌细胞凋亡的作用。 CaSR的激活下调Bcl-2表达,上调caspase-3和Fas / FasL表达并刺激ERK1 / 2磷酸化。总之,CaSR通过抑制Bcl-2,诱导钙超载和激活Fas / FasL死亡受体途径,参与新生大鼠心室心肌细胞的I / R损伤和凋亡。

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