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.
出处
《病毒学报》
CAS
CSCD
北大核心
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
