畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (7): 1201-1207.doi: 10.11843/j.issn.0366-6964.2015.07.017

• 预防兽医 • 上一篇    下一篇

细胞自噬对牛病毒性腹泻病毒复制的影响

宫晓炜,郑福英,陈启伟,刘永生*,李兆才,周继章*,才学鹏,殷宏   

  1. (中国农业科学院 兰州兽医研究所 家畜疫病病原生物学国家重点实验室 草食动物细菌病创新团队,兰州 730046)
  • 收稿日期:2014-10-10 出版日期:2015-07-23 发布日期:2015-07-23
  • 通讯作者: 刘永生,主要从事细菌病致病机制及防控技术研究,E-mail:liuyongsheng@caas.cn;周继章,主要从事动物疫苗及分子免疫学研究,E-mail:zhoujizhang@126.com
  • 作者简介:宫晓炜(1982-),女,甘肃兰州人,助理研究员,博士,主要从事动物疫苗与分子免疫学研究,Tel:0931-8342673,E-mail:gongxiaowei@caas.cn
  • 基金资助:

    国家肉牛产业技术体系项目(CARS-38)

Impact of Cellular Autophagy on Bovine Viral Diarrhea Virus Replication

GONG Xiao-wei,ZHENG Fu-ying,CHEN Qi-wei,LIU Yong-sheng*,LI Zhao-cai,ZHOU Ji-zhang*,CAI Xue-peng,YIN Hong   

  1. (State Key Laboratory of Veterinary Etiological Biology,Bacterial Disease in Grazing Animal Team,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)
  • Received:2014-10-10 Online:2015-07-23 Published:2015-07-23

摘要:

本研究旨在了解牛病毒性腹泻病毒(BVDV)诱导的细胞自噬在病毒感染过程中所起的作用。用Oregon C24V株感染MDBK细胞,以Western blot、间接免疫荧光、透射电镜三种方法鉴定细胞自噬的产生;通过自噬抑制药物Wortmannin和自噬基因Beclin-1的RNAi干扰阻断自噬,探究自噬对BVDV复制的影响。结果显示,病毒感染可以引起LC3Ⅰ向LC3Ⅱ的转化及p62的降解;转染GFP-LC3质粒的细胞,在病毒感染后出现增多的聚集颗粒;透射电镜能观察到细胞中出现大量的双层膜结构;此外,抑制细胞自噬或干扰Beclin-1的表达,细胞上清中病毒的滴度及病毒蛋白质的表达量都有所降低。BVDV感染早期可以诱导自噬的产生,且自噬对病毒的复制有利。

Abstract:

The objective of this study was to investigate the role of cellular autophagy in bovine viral diarrhea virus (BVDV) infection.We used Oregon C24V strain to determine whether BVDV infection induced autophagy by using western blot,indirect immunofluorescent assay and transmission electron microscopy.To analyze the effect of autophagy on the replication of BVDV in MDBK cell,wortmannin and small interfering RNAs targeting Beclin-1 were used to inhibit autophagy.The results showed that BVDV infection results in LC3-Ⅰ/Ⅱ conversion,p62 degradation,an increased accumulation of punctate GFP-LC3-expressing cells,and a higher number of autophagosome-like double-membrane vesicles in the cytoplasm of host cells.Inhibition of autophagy or interfering expression of Beclin-1 led to a significant reduction in BVDV titers and protein expression.These results demonstrate that BVDV infection induces autophagy which in turn enhances BVDV replication.

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