【摘要】 目的:探讨活化的钙敏感性受体与心肌细胞缺血/再灌注损伤凋亡是否有关。方法:新生鼠心肌细胞在模拟心肌缺血状态下,培养2 h后,在标准培养液中再培养24 h,从而建立一个模拟心肌缺血/再灌注模型。使用末端脱氧核苷酰基转移酶介导性dUTP切口末端标记(TUNEL),检测心肌细胞凋亡。钙敏感性受体mRNA表达通过逆转录聚合酶链式反应(RT-PCR)测定。采用免疫蛋白印迹法(Western blotting),测定Caspase-3和Bcl-2。结果:模拟I/R增强了钙敏感性受体表达及心肌细胞凋亡。GdCl3,一种特殊的CaSR激活剂进一步增强了钙敏感性受体表达及心肌细胞凋亡,而对Caspase-3和Bcl-2则分别起着正向及负向调节作用。结论:CaSR与新生鼠心肌细胞缺血/再灌注损伤及心肌细胞凋亡有关,其促进了Caspase-3而抑制了Bcl-2的表达。更多还原

【Abstract】 Objective:To examine the association of activation of calcium-sensing receptors(CaSR) with apoptosis in cardiomyocytes under simulated ischemia/reperfusion. Methods:Ventricular cardiomyocytes of neonatal rats were incubated in ischemia-mimetic solution for 2 h,then re-incubated in normal culture medium for 24 h to establish a model of simulated ischemia/reperfusion(I/R).Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL assay).The expression of CaSR mRNA was detected by reverse transcriptase polymerase chain reaction(RT-PCR).The expression of Caspase-3 and Bcl-2 was detected by Western blotting. Results:The simulated I/R enhanced the expression of CaSR and cardiomyocyte apoptosis.GdCl3,a specific activator of CaSR,further increased the expression of CaSR and cardiomyocyte apoptosis,along with upregulation of Caspase-3 and downregulation of Bcl-2. Conclusion:CaSR is associated with I/R injury and apoptosis in neonatal rat ventricular cardiomyocytes via suppressing Bcl-2 and promoting Caspase-3 expression.更多还原