猪流行性腹泻病毒感染Vero-E6细胞诱导凋亡机制的研究

Induction of apoptosis in Vero-E6 cells infected with porcine epidemic diarrhea virus

为研究猪流行性腹泻病毒(PEDV)诱导体外细胞凋亡的机理,本研究利用荧光定量PCR方法结合流式细胞术检测PEDV感染Vero-E6细胞后凋亡分子的变化情况。荧光定量PCR结果表明,抑制细胞凋亡因子Bcl-2在PEDV感染后基因转录水平迅速升高,在24 h达到峰值后被抑制;而促凋亡因子的蛋白水解酶caspase8和caspase3在PEDV感染后基因转录水平也随之提高,在48 h达到峰值。流式细胞术结果表明PEDV感染细胞48 h后促凋亡分子中的蛋白水解酶Caspases被活化。该研究结果揭示了PEDV感染细胞能够拮抗细胞存活信号通路,引起细胞凋亡,其中凋亡调节因子Bcl-2、caspase3和caspase8在细胞凋亡发生过程中起关键作用。

Porcine epidemic diarrhea virus （PEDV） causes a severe infectious enteric disease and has triggered great economic losses in swine industry.To study the mechanism of virus-induced apoptosis,Vero-E6 cells were infected with PEDV and the mRNA expressions of the apoptotic factors：Bcl-2,caspase3 and caspase8 were detected by real-time PCR.The results showed that these three apoptotic factors were up-regulated during virus infection; the anti-apoptotic factor Bcl-2 reached the peak at 24 hours post-infection; and the pro-apoptotic factors caspase3 and caspase8 reached a maximum at 48 hours post-infection.In addition,the pro-apoptotic factors of caspases were activated at 48 hours post-infection detected by flow cytometry.These findings demonstrated that PEDV infection was able to block anti-apoptotic signal pathway and induce cell apoptosis,which indicates that the apoptotic factors Bcl-2,caspase3 and caspase8 play a key role in the process of cell apoptosis.