细胞病变型牛病毒性腹泻病毒对宿主细胞自噬作用的研究  

Research of Attenuated Bovine Viral Diarrhea Virus Effecting on Host Cell Autophagy

作　　者：李家伟[1] 李健友[2] 王超[3] 张淑琴[3] 程世鹏[3] 郭利[3] 

机构地区：[1]吉林农业大学研究生院,长春130118 [2]东北农业大学,哈尔滨150030 [3]中国农业科学院特产研究所特种动物分子生物学重点实验室,长春130112

出　　处：《病毒学报》2017年第3期435-440,共6页Chinese Journal of Virology

基　　金：国家自然科学基金(项目号:31302097),题目:自噬在牛病毒性腹泻病毒不同生物型感染宿主细胞中的作用及相关基因表达研究~~

摘　　要：细胞自噬(Autophagy)是一种真核生物细胞内损伤的细胞器和长寿命蛋白通过双层膜结构的自噬小泡包裹后,送入溶酶体或液泡中进行降解并得以循环利用的高度保守的分解代谢过程。本研究旨在了解细胞病变型(CEP)牛病毒性腹泻病毒(BVDV NM)对宿主细胞MDBK细胞自噬作用的影响。实验使用BVDV NM株感染宿主细胞MDBK,在不同时间点分别通过透射电镜观察自噬体形成、报告荧光GFP-LC3自噬底物检测以及Western blot方法鉴定细胞自噬标记物LC3-Ⅰ/LC3-Ⅱ和P62的表达等试验方法对自噬进行检测。结果显示,感染BVDV NM株的MDBK细胞出现了明显的细胞病变;透射电镜能观察到细胞中存在大量的双层膜结构的自噬泡;转染GFP-LC3荧光质粒后,在感染病毒的细胞内出现增多的聚集绿色荧光自噬小体;此外,随着BVDV NM株感染MDBK时间的延长可以发现LC3-Ⅰ/Ⅱ蛋白的量逐渐增加以及P62蛋白的降解。试验表明BVDV NM株感染MDBK可以促进细胞自噬的发生。Autophagy is a highly conserved catabolic process in eukaryotes in which the damaged organelles and long lived proteins are packaged by a double membraned vesicle known as an autophagosome and then transmitted into lysosome or vacuole to be degraded and recycled. The obiective of this study was to investigate the relationship between autophagy and replication of bovine viral diarrhea virus （BVDV）. We used a BVDV strain （NM） to infect its host cell MDBK and determined whether this infection induced autophagy by observing cytopathy, transmission electron microscopy, transfecting fluorescence plasmid GFP-LC3 and Western blot. The results revealed that MDBK infected by BVDV showed cytopathy and a higher number of autophagosome-like vesicles with double membrane were observed by transmission electron microscopy. After transfecting fluorescence plasmid GFP-LC3, the number of green fluorescent proteins increased in infected cells. With extension in the infection time of MDBK by BVDV strain, LC3-II gradually increased while p62 degraded. The results indicated that infecting MDBK by BVDV NM strain can induce autophagy.

关 键 词：牛病毒性腹泻病毒(BVDV) 细胞自噬 

分 类 号：R373.1[医药卫生—病原生物学]