番鸭呼肠孤病毒感染诱导细胞自噬的研究  

Studies on Cellular Autophagy Induced by the Muscovy Duck Reovirus

作　　者：吴异健[1,2] 崔龙萍[1] 黄诗杭 王全溪[1,2] 吴晓平[1,2] 周五朵 朱二鹏 黄一帆[1,2] 吴宝成[1,2] 

机构地区：[1]福建农林大学动物科学学院,福州350002 [2]福建省兽医中药与动物保健重点实验室,福州350002

出　　处：《病毒学报》2017年第2期205-210,共6页Chinese Journal of Virology

基　　金：国家自然科学基金;项目编号(u1305212);题目:闽台特色水禽重要病毒的致病机理研究;国家自然科学基金;项目编号(31372474);题目:猴头菇多糖对MDRV致番鸭肠黏膜免疫功能损伤的修复及其机制研究

摘　　要：本研究通过透射电镜观察自噬体的形态学结构,蛋白免疫印迹(Western blot)检测细胞自噬相关蛋白LC3II和磷酸化mTOR(p-mTOR)表达量的动态变化趋势,探索番鸭呼肠孤病毒(MDRV)感染对细胞自噬的影响。结果显示,MDRV-YB株感染的DF-1细胞中可见典型的包裹胞浆成分的双层膜结构;MDRV感染番鸭成纤维细胞(MDFs)和DF-1细胞后0~60h内,其LC3II表达量均呈现先升高后降低的趋势,且均在感染后36h达到最大值,其p-mTOR表达量均呈现先降低后回升的趋势,且分别在感染后36h和24h达到最低值;灭活MDRV组细胞未见LC3II表达。结果表明MDRV能够诱导感染的MDFs和DF-1细胞持续较长时间细胞自噬,而灭活的MDRV不能引起细胞自噬。To study the effects of the Muscovy duck reovirus（MDRV）on cellular autophagy,an autophagosome-like structure was observed by transmission electron micro scopy（TEM）,and expression of the autophagy-related proteins LC3 II and phospho-mTOR were detected by western blotting.TEM showed typical bilayer membrane structures containing cytoplasmic inclusions to be visible in MDRV-YB-infected DF-1cell lines.Western blotting showed that LC3 II expression was sustained within 60 hours post-infection（hpi）.LC3 II expression displayed a significant increase and subsequent decrease in the number of primary Muscovy duck fibroblasts（MDFs）and DF-1cells infected with the MDRV-YB strain,with an identical maximum at 36 hpi.Unsurprisingly,phospho-mTOR expression decreased and then rebounded,whereas a minimum was obtained at 36 hpi and 24 hpi,respectively.Inactivated MDRV did not induce LC3 II expression.These results showed that MDRV could trigger autophagy of relatively long duration in primary MDFs and DF-1cells,and that inactivated MDRV did not induce autophagy.

关 键 词：番鸭呼肠孤病毒 细胞自噬 LC3Ⅱ 蛋白免疫印迹(Western blot) 透射电镜 

分 类 号：S852.65[农业科学—基础兽医学]