畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (9): 1481-1487.doi: 10.11843/j.issn.0366-6964.2014.09.014

• 预防兽医 • 上一篇    下一篇

猪瘟病毒促进细胞自噬并利于病毒增殖

康恺,林鸷,高海慧,张成成,李河林,梁武龙,王静,曹志,张彦明*   

  1. (西北农林科技大学 动物医学院,杨凌 712100)
  • 收稿日期:2014-02-27 出版日期:2014-09-23 发布日期:2014-09-23
  • 通讯作者: 张彦明(1956-),E-mail: ylzhangym@sohu.com
  • 作者简介:康恺(1986-),女,陕西岐山人,博士,主要从事分子病原学与免疫学研究,E-mail: kaikang0@163.com
  • 基金资助:

    国家自然科学基金项目(31172339)

Classical Swine Fever Virus Promotes Cell Autophagy which Facilitates Virus Proliferation

KANG Kai,LIN Zhi,GAO Hai-hui,ZHANG Cheng-cheng,LI He-lin,LIANG Wu-long,WANG Jing,CAO Zhi,ZHANG Yan-ming*   

  1. (College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China)
  • Received:2014-02-27 Online:2014-09-23 Published:2014-09-23

摘要:

细胞自噬在病毒的增殖、释放中有着重要的作用,目前国内仍然没有猪瘟病毒与细胞自噬相互作用关系的报道。本研究旨在探讨猪瘟病毒Shimen株感染宿主细胞(ST)后对细胞自噬的影响,以及细胞自噬对病毒增殖的作用。通过透射电镜和自噬体标记分子LC3的荧光聚点的观察,直观判断病毒对自噬的影响;并通过Western blot,检测LC3蛋白和P62蛋白的表达量,以及LC3蛋白转化分析,确定猪瘟病毒感染促进细胞自噬发生。利用自噬促进剂雷帕霉素、抑制剂3-MA、自噬体与溶酶体融合阻断剂氯喹(CQ),以及自噬基因Beclin 1、LC3蛋白的干扰,调控自噬来研究自噬对病毒增殖的影响。结果表明,病毒感染细胞促进了细胞自噬的发生,且能形成完整的自噬过程,同时病毒利用细胞自噬来促进自身增殖。本研究为探索猪瘟病毒与宿主细胞相互作用关系增加了新的数据。

Abstract:

Autophagy plays an important role in virus proliferation and release while there have no report about the relationship of classical swine fever virus (CSFV) and host cell autophagy.This study was undertaken to elucidate the impact of CSFV Shimen strain on host cell (ST) autophagy and the effect of autophagy to virus proliferation.The transmission electron microscopy (sem),LC3 dots,biomarkers of autophagy,were adopt to get a preliminary determination.The protein expression of LC3 and P62,as well as conversion of LC3Ⅰto LC3 Ⅱwere semi-quantified by Western blot,the results suggested that the CSFV promotes cell autophagy.Autophagy promoter rapamycin,inhibitors 3-MA,chloroquine(CQ),fusion blocker of autophagosome and lysosomes,were used to regulate the autophagy,as well as autophagy associated gene (ATG) Beclin 1 and LC3 protein were interfered to investigate the effect of autophagy on virus proliferation.The results showed that virus infection promotes cell autophagy and form complete autophagy flux,which facilitates virus proliferation.This research provides new information to the relationship of CSFV and host cell.

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