mTOR信号通路在调控猪繁殖与呼吸综合征病毒复制中的作用  

The role of mTOR signaling pathway in regulating porcine reproductive and respiratory syndrome virus replication

作　　者：李华玮[1,2] 赵绪永 赵孟孟[3] 李新锋 姬向波 乔松林 LI Huawei;ZHAO Xuyong;ZHAO Mengmeng;LI Xinfeng;JI Xiangbo;QIAO Songlin(College of Food and Bioengineering,Henan University of Animal Husbandry and Economics,Zhengzhou 450046,China;Henan Key Laboratory of Innovation and Utilization of Unconventional Feed Resources, Zhengzhou 450046,China;School of Life Science and Engineering,Foshan University,Foshan 528000,China;Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

机构地区：[1]河南牧业经济学院食品与生物工程学院,河南郑州450046 [2]河南省非常规饲料资源创新利用重点实验室,河南郑州450046 [3]佛山科学技术学院生命科学与工程学院,广东佛山528000 [4]河南省农业科学院动物免疫学重点实验室,河南郑州450002

出　　处：《河南农业大学学报》2022年第1期106-114,共9页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(31902284,31902279);河南省高等学校青年骨干教师培养计划(2020GGJS258);河南牧业经济学院博士科研启动基金项目(2019HNUAHEDF040);河南牧业经济学院科技创新团队(2018KYTD13)。

摘　　要：【目的】为阐明哺乳动物雷帕霉素靶点(mammalian target of Rapamycin,mTOR)信号通路在调控猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)复制过程中的作用,探索防控该病传染的新途径。【方法】PRRSV感染猪肺泡巨噬细胞(porcine alveolar macrophage,PAMs)24 h后,用蛋白质印迹法检测mTOR信号通路下游信号分子的表达水平及磷酸化水平。【结果】在PRRSV感染PAMs后,mTOR、eIF4E结合蛋白1(eukaryotic initiation factor 4E binding protein 1,4EBP1)及PS6K的磷酸化水平显著增加,表明PRRSV感染激活了mTOR信号通路。用mTOR特异性抑制剂雷帕霉素(Rapamycin)处理PAMs,发现Rapamycin能够显著抑制PRRSV感染引起的mTOR信号通路下游关键信号分子4EBP1磷酸化,且剂量依赖性和时间依赖性促进了PRRSV的复制。【结论】mTOR信号通路参与调控PRRSV的复制,Rapamycin对4EBP1磷酸化有抑制作用,对PRRSV的复制有促进作用。该研究为后续进一步探索PRRSV复制的翻译起始机制奠定基础。【Objective】To clarify the role of mammalian target of Rapamycin(mTOR)pathway in regulating the replication of porcine reproductive and respiratory syndrome virus(PRRSV);To explore new ways to prevent and control porcine reproductive and respiratory syndrome(PRRS).【Methods】After PRRSV infected PAMs for 24 h,Western Blot was used to detect the expressional level and phosphorylation level of signal molecules downstream of mTOR signaling pathway.【Results】After PAMs was infected with PRRSV,the phosphorylation levels of mTOR,4EBP1 and PS6K increased significantly,indicating that PRRSV infection activated mTOR signaling pathway.After PAMs was treated with Rapamycin,a specific inhibitor of mTOR,it was found that Rapamycin could significantly inhibit the phosphorylation of 4EBP1 which caused by PRRSV infection,and promote the replication of PRRSV in a dose-dependent and time-dependent manner.【Conclusion】mTOR signaling pathway involved in the regulation of PRRSV replication.Rapamycin can inhibit phosphorylation level of P-4EBP1 and promote PRRSV replication.This study has laid the foundation for further exploring the translation initiation mechanism of PRRSV replication.

关 键 词：猪繁殖与呼吸综合征病毒 哺乳动物雷帕霉素靶点 信号通路 猪肺泡巨噬细胞 蛋白质印迹法 病毒复制 

分 类 号：S855.3[农业科学—临床兽医学]