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目的 探讨细胞内游离钙 [Ca2 + ]i在柯萨奇病毒 B3(CVB3)诱导培养心肌细胞凋亡中的作用。方法 DNA裂点检测法 (3′-末端标记 )及透射电镜检测细胞凋亡。 Fluo3- AM负载心肌细胞 ,共聚焦显微镜观察[Ca2 + ]i荧光强度变化。结果 感染 2 4h心肌细胞内 CVB3滴度达峰值。感染 10 h未见凋亡的心肌细胞 ,17、2 4和 36 h凋亡细胞分别为 5 %、6 0 %和 90 % ,感染 17h心肌 [Ca2 + ]i浓度达峰值。电镜结果提示 CVB3感染组存在典型的凋亡心肌细胞。结论 CVB3可诱导培养心肌细胞的凋亡过程。细胞内 Ca2 + 参与凋亡细胞的信息传导过程 ,并在凋亡早期发挥重要作用
Objective To investigate the role of intracellular free calcium [Ca2 +] i in cardiomyocyte apoptosis induced by coxsackievirus B3 (CVB3). Methods Cell apoptosis was detected by DNA dehiscence assay (3’-end labeling) and transmission electron microscopy. Fluo3-AM loaded cardiomyocytes were observed by confocal microscopy [Ca2 +] i fluorescence intensity changes. RESULTS: The intracellular CVB3 titer peaked 24 h after infection. No apoptotic cardiomyocytes were found at 10 h after infection, and apoptotic cells were 5%, 60% and 90% respectively at 17, 24, and 36 h after infection, and the peak of [Ca2 +] i concentration was observed at 17 h after infection. Electron microscopy showed that there were typical apoptotic cardiomyocytes in CVB3 infection group. Conclusion CVB3 can induce cardiomyocyte apoptosis. Intracellular Ca2 + is involved in the process of information transduction of apoptotic cells and plays an important role in the early stages of apoptosis