H9N2流感病毒诱导MDCK细胞自噬对病毒复制的影响  

The effects of autophagy induced by H9N2 influenza virus on virus replication in MDCK cells

作　　者：李春红[1] 王丽[1] 董玉龙[2] 李欣蕊 杨昊 魏鑫 徐彤[1] LI Chunhong;WANG Li;DONG Yulong;LI Xinrui;YANG Hao;WEI Xin;XU Tong(Key Laboratory of Preventive Veterinary Medicine,Department of Veterinary Medicine,Animal Science College,Hebei North University,Zhangjiakou,Hebei Province 075000,China;College of Science,Hebei North University,Zhangjiakou,Hebei Province 075000,China)

机构地区：[1]河北北方学院预防兽医学重点实验室,河北张家口075000 [2]河北北方学院理学院,河北张家口075000

出　　处：《中国饲料》2022年第17期12-16,23,共6页China Feed

基　　金：河北省第二期现代农业产业技术体系蛋肉鸡创新团队建设专项经费(HBCT2018150207);河北省科技厅重点研发计划(20326624D);河北省高等学校科学技术研究项目(ZD2020314);河北省高层次人才资助项目(A202003006);河北北方学院校级科研项目(YB2020006,XJ2021008);河北北方学院校级教改项目(GJ2020241)。

摘　　要：本试验旨在研究H9N2流感病毒感染MDCK细胞引发的细胞自噬对病毒复制的影响。试验首先建立了稳定表达GFP-LC3的MDCK细胞系,该细胞系经H9N2流感病毒感染后,激光共聚焦观察到了绿色荧光的点状聚集,得出H9N2流感病毒感染MDCK细胞引发细胞自噬。随后试验利用3-MA抑制细胞自噬、Rapamycin诱导细胞自噬,荧光定量PCR检测NP mRNA水平,TCID50检测病毒的滴度进行细胞自噬与病毒复制的研究。结果表明:Rapamycin可显著增加NP mRNA水平和病毒滴度(P <0.01);3-MA可以显著降低NP m RNA水平和病毒滴度(P<0.01)。说明H9N2流感病毒感染MDCK细胞引发的细胞自噬可促进流感病毒的复制。To investigate the effects of autophagy induced by H9N2 influenza virus on virus replication in MDCK cells,firstly MDCK cells line with expression of GFP-LC3 was constructed.After the cells line was infected by H9N2 influenza virus,spot-like aggregati on of green fluorescence was observed by confocal laser scanning.Autophagy was caused by H9N2 influenza virus in MDCK cells in this experiment.Besides,to study whether autophagy affects virus replication,3-MA was used to inhibit autophagy,and then Rapamycin was used to cause autophagy,so the mRNA level of NP was detected by real-time PCR and the titer of virus was detected by TCID50.The results showed that 3-MA reduced the NP mRNA levels and the virus titer,moreover,Rapamycin was contrary to the action of 3-MA and increased the two indexes.The conclusion of this study showed that autophagy induced by H9N2 influenza virus infection in MDCK cells could promote influenza virus replication.

关 键 词：H9N2流感病毒 细胞自噬 病毒复制 MDCK细胞 诱导 

分 类 号：S811.6[农业科学—畜牧学]