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为研究大鲵蛙病毒(Chinese giant salam ander ranavirus,CGSRV)对细胞凋亡的影响,以CGSRV感染鲤鱼上皮瘤(EPC)细胞,分别于感染后第0、3、6、9、12、15小时收集细胞,用倒置显微镜和透射电镜观察感染的EPC细胞的形态学变化,Annexin V-FITC/PI双染与JC-1染色后用流式细胞仪检测细胞凋亡率与线粒体膜电位(Δψm)崩解率。结果显示,EPC细胞在感染后第6小时出现细胞收缩变圆,脱落,漂浮于培养基中;电镜下,被感染的EPC细胞的细胞核出现典型的染色质边集呈月牙形和染色质浓缩形成凋亡小体的现象。流式细胞仪检测结果显示,感染后第3小时即出现凋亡率和线粒体膜电位崩解率显著上升,6 h后实验组各时间点凋亡率和线粒体膜电位崩解率都极显著地高于对照组(P<0.01)。结果表明,CGSRV感染EPC细胞可通过线粒体途径诱导EPC细胞凋亡。
To investigate the effect of CGSRV on apoptosis, CGSRV was used to infect carp epithelial tumor (EPC) cells and were collected at 0, 3, 6, 9, 12 and 15 hours after infection The morphological changes of infected EPCs were observed by inverted microscope and transmission electron microscope. The apoptotic rate and mitochondrial membrane potential (Δψm) collapse were detected by flow cytometry after Annexin V-FITC / PI double staining and JC-1 staining Solution rate. The results showed that EPCs became round at the 6th hour after infection, shedding and floating in the medium. Under the electron microscope, typical chromatin margins appeared in the nucleus of infected EPCs in crescent and chromatin condensation Apoptosis phenomenon. Flow cytometry results showed that the apoptotic rate and mitochondrial membrane potential disintegration rate increased significantly at the 3rd hour after infection, and the apoptotic rate and the mitochondrial membrane potential disintegration rate of the experimental group at 6 hours were significantly Higher than the control group (P <0.01). The results showed that EPCs infected with CGSRV could induce EPCs apoptosis through mitochondria pathway.