猪流行性腹泻病毒诱导Vero E6细胞凋亡的动态研究

Dynamic analysis of apoptosis in Vero E6 cells induced by porcine epidemic diarrhea virus

为研究猪流行性腹泻病毒(PEDV)诱导体外细胞产生病变及细胞凋亡的动态变化,本研究应用western blot、间接免疫荧光、细胞活性计数(CCK-8)等方法检测PEDV感染Vero E6细胞后不同时间点细胞内病毒含量、细胞活性、凋亡小体的形成,以及相关凋亡因子的变化。结果表明,感染后24 h细胞内病毒含量开始明显升高,48 h时达到峰值。同时在病毒感染24 h后出现明显的细胞病变,感染48 h后出现典型的凋亡特征,细胞聚堆,细胞核内染色质固缩,形成凋亡小体。CCK-8分析检测结果显示,与对照组相比感染病毒细胞活性明显降低。另外,western blot结果表明促凋亡因子蛋白水解酶Caspase-3在病毒感染过程中被活化。该研究结果进一步证实了PEDV感染可以诱导细胞凋亡,其中Caspase-3的激活在病毒诱导细胞凋亡的过程中发挥重要作用。

To investigate the apoptosis in Vero E6 cells infected with porcine epidemic diarrhea virus （PEDV）, western blot, indirect immunofluorescence assay, Cell Counting Kit-8 （CCK-8） were used to detect virus load, cell activity, the formation of apoptotic bodies and the variation of apoptosis factors at different times. The results indicated that the PEDV increased significantly at 24 hrs and reached at peak at 48 hrs post infection. We could observe obvious cytopathic effects at 24 hrs and typical apoptotic features including chromatin condensation, nuclear fragmentation and apoptotic bodies at 48 hrs post infection. CCK-8 assay reflected that the activity of cells infected with PEDV decreased compared with normal cells. In addition, the pro-apoptotic factor of Caspase-3 was activated induced by PEDV infection through western blot assay. These findings further verified that PEDV infection could induce cell apoptosis and the activation of Caspase-3 played important role in cell apoptosis induced by PEDV infection.