摘要
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。
In order to detect the character of apoptosis induced by Chinese giant salamander ranavirus（CGSRV）,a fish cell line,epithelioma papillosum cyprini（EPC） cells,was infected by CGSRV.The EPC was collected at hour 0,3,6,9,12 and 15 post-infection to observe the alteration of morphology by inverted microscope and transmission electron microscope,and to detect the apoptotic percentage and the mitochondrion membrane potential（Δψm） decreased ratio by flowcytometry after Annexin V-FITC/PI dyeing and JC-1 dyeing,respectively.The results showed that the cells changed to round and detachment off the plate at hour 6 post-infection;and some typical apoptotic characteristics,such as nuclear chromatin concentration and edge accumulation,were observed by transmission electron microscope.Furthermore,the apoptotic percentage and the mitochondrion membrane potential decreased ratio were significanthly higher than that of the control group at hour 3 post-infection and extremely significant higher than that of the control group at the following time（P〈0.01）.The results revealed the mitochondrion-mediated apoptosis induced by CGSRV infection in EPC.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第8期1062-1066,共5页
Chinese Veterinary Science
基金
四川省科技支撑计划项目(2014NZ0027)
四川省淡水鱼产业技术体系创新团队建设项目
关键词
大鲵蛙病毒
鲤鱼上皮瘤细胞
细胞凋亡
线粒体膜电位
Chinese giant salamander ranavirus（CGSRV）
epithelioma papillosum cyprini（EPC）
apoptosis
mitochondrial membrane potential
