牛病毒性腹泻病毒诱导铁死亡对其复制水平的影响  

The Effects of Bovine Viral Diarrhoea Virus(BVDV)-induced Ferroptosis on Virus Replication

作　　者：张梓璇 张颖[1,2] 李志军 杨婧玲[1,2] 蒋子豪 黄华敏 齐雪峰 ZHANG Zixuan;ZHANG Ying;LI Zhijun;YANG Jingling;JIANG Zihao;HUANG Huamin;QI Xuefeng(College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China;Key Laboratory of Ruminant Disease Prevention and Control(West),Ministry of Agriculture and Rural Affairs,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物医学院,杨凌712100 [2]农业农村部反刍动物重大疫病防控重点实验室(西部),杨凌712100

出　　处：《畜牧兽医学报》2024年第12期5716-5724,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家级大学生创新创业训练计划项目(202310712186);陕西省农业科技创新计划项目(NYKJ-2022-YL(XN)08)。

摘　　要：本研究旨在探究牛病毒性腹泻病毒(BVDV)感染对牛肾细胞(Madin-Darby Bovine Kidney cells,MDBK)铁死亡的调控作用及其对病毒复制的影响。本研究通过激光共聚焦、Western blot、qPCR和电镜技术等技术检测BVDV感染MDBK细胞铁死亡发生与病毒复制水平,另外,使用铁死亡抑制剂Fer-1研究铁死亡对病毒复制和细胞炎性因子表达的影响。结果表明:BVDV(MOI=5)感染细胞较正常对照组细胞死亡率显著增加(P<0.05),感染细胞内Fe^(2+)和脂质过氧化水平较正常细胞显著升高(P<0.05),透射电镜检测可见线粒体表面积变小、嵴减少以及膜密度增加等铁死亡发生特征;将铁死亡抑制剂Fer-1预处理MDBK细胞后,可显著抑制BVDV感染病毒复制水平(P<0.05);此外,BVDV感染诱导铁死亡对MDBK细胞中IL-1β、IL-18以及IFN-β等炎性因子呈正调控效应。以上结果提示,BVDV感染可引起宿主细胞铁死亡,且铁死亡发生可显著促进病毒复制水平以及宿主细胞炎性因子表达。This study aimed to investigate the regulatory effect of bovine viral diarrhoea virus(BVDV)infection on ferroptosis in Madin-Darby Bovine Kidney(MDBK)cells and its effect on viral replication.Technologies such as confocal microscopy,Western blot,qPCR and electron microscopy were used to detect ferroptosis and the level of viral replication in BVDV-infected cells.Furthermore,the viral replication level and inflammatory cytokines expression in BVDV-infected cells pretreated with Fer-1,a commonly used ferroptosis inhibitor,were also detected.The results showed that,compared with the normal control group,the mortality rate of cells infected with BVDV(MOI=5)was significantly increased(P<0.05),which was accompanied with increased levels of Fe^(2+)and lipid peroxidation(P<0.05).Transmission electron microscopy(TEM)analysis revealed that BVDV-infected and Erastin-treated cells displayed shrunk mitochondria with fewer cristae,increased mitochondrial membrane density and decreased mitochondrial mean areas compared with those of mock-infected cells,which was a typical morphological feature of ferroptosis.Fer-1 treatment significantly inhibited the levels of viral replication in BVDV-infected cells(P<0.05).In addition,ferroptosis induced by BVDV infection had positive regulatory effects on inflammatory cytokines expression,including IL-1β,IL-18 and IFN-β.The results suggested that BVDV infection can induce ferroptosis in host cells,and the induction of ferroptosis enhanced the level of viral replication and the expression of inflammatory cytokines.

关 键 词：牛病毒性腹泻病毒(BVDV) 铁死亡 病毒复制 炎性因子 

分 类 号：S852.653[农业科学—基础兽医学]