鹅细小病毒诱导番鸭胚成纤细胞自噬  

Autophagy induced by goose parvovirus in Muscovy duck embryo fibroblasts(MDEFs)

作　　者：王劭[1] 程晓霞[1] 肖世峰 林锋强[1] 朱小丽[1] 陈少莺[1] 陈仕龙[1] WANG Shao;CHENG Xiaoxia;XIAO Shifeng;LIN Fengqiang;ZHU Xiaoli;CHEN Shaoying;CHEN Shilong(Fujian Animal Diseases Control Technology Development Center,Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《中国兽医学报》2023年第8期1643-1650,共8页Chinese Journal of Veterinary Science

基　　金：福建省自然科学基金资助项目(2021J01482);福建省农业科学院农业科技专项资助项目(GJPY2019004);福建省农业科学院创新团队资助项目(CXTD2021034);福建省公益类科研院所专项资助项目(2021R10260010,2022R1026004)。

摘　　要：细胞自噬在病毒生命周期中发挥着重要作用,目前国内仍然没有鹅细小病毒(GPV)与细胞自噬关系的系统研究。本研究旨在探讨番鸭小鹅瘟病毒(MDGPV)PT株与短喙矮小综合征鹅细小病毒(SBDS-GPV)M15株细胞适应毒感染番鸭胚成纤维细胞(MDEFs)后对细胞自噬的影响,以及细胞自噬对病毒增殖的作用。通过激光共聚焦方法观察自噬标记物LC3-Ⅱ特异绿色荧光聚集颗粒,借助蛋白免疫印迹检测LC3蛋白与p62蛋白的表达量,以及LC3-Ⅰ/LC3-Ⅱ蛋白转化分析,确定GPV感染能够促进细胞自噬发生。利用膜通透性半胱氨酸蛋白酶抑制剂阿洛司他丁、自噬诱导剂雷帕霉素以及自噬抑制剂3-甲基腺嘌呤,调控自噬来研究细胞自噬对病毒增殖的影响,结果显示细胞自噬有利于GPV在MDEFs中的复制。结果表明GPV感染能够诱导细胞自噬,同时细胞自噬有利于病毒在宿主细胞中的复制。Autophagy plays an important role in the virus life cycle.At present,there is still no systematic study on the relationship between goose parvovirus and autophagy in China.This study was undertaken to elucidate the impact of cell-derived GPV strains(Muscovy duck-origin goose parvovirus,MDGPV-PT and short beak and dwarfism syndrome virus,SBDS-GPV M15)on host cells(Muscovy duck embryo fibroblasts,MDEFs)autophagy and the effect of cell autophagy to virus proliferation.In the present study,we showed that GPV infection markedly increased the autophagy marker LC3-Ⅱspecific green fluorescent aggregation particles in the cytoplasm of MDEFs by the confocal fluorescence microscopy.The protein expression of LC3 and p62,as well as LC3-Ⅰ/LC3-Ⅱconversion were detected by Western blot,the results suggested that GPV promotes cell autophagy.The membrane-permeable cysteine protease inhibitor aloxistatin(E64d),autophagy promoter rapamycin(RAPA),and autophagy inhibitor 3-methyladenine(3-MA)were respectively used to regulate the autophagy and to investigated the effect of autophagy on GPV proliferation.The results indicated that the autophagy benefited the GPV replication in MDEFs.Collectively,GPV induces autophagy in host cells,and the autophagy promotes the replication of GPV.

关 键 词：鹅细小病毒 细胞自噬 病毒复制 LC3 P62 

分 类 号：S852.65[农业科学—基础兽医学]