猪伪狂犬病病毒诱导猪肺泡巨噬细胞氧化应激和炎症反应研究  

Oxidative Stress and Inflammatoin Induced by Pseudorabies Virus Infection in 3D4/2 Cells

作　　者：于美玲 缪卫媛 吴苑滢 梁颖 曹迷霞 韦英益[1] 王蕾 胡庭俊[1] YU Mei-ling;MIAO Wei-yuan;WU Yuan-ying;LIANG Ying;CAO Mi-xia;WEI Ying-yi;WANG Lei;HU Ting-jun(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi,530005,China;Nanning Institute for Food and Drug Control,Nanning,Guangxi,530005,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530005 [2]南宁市食品药品检验所,广西南宁530005

出　　处：《动物医学进展》2022年第6期13-21,共9页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32072907);广西自然科学基金青年基金项目(2019JJB130197)。

摘　　要：旨在通过猪伪狂犬病病毒(PRV)感染猪肺泡巨噬细胞(3D4/2),观察PRV感染对猪免疫细胞氧化应激和炎性反应的影响,为进一步研究PRV感染致病机制和防控药物的开发奠定基础。采用猪肾脏细胞(PK-15)扩增PRV,测定PRV TCID_(50),观察PRV感染3D4/2细胞病变及检测病毒扩增情况,测定PRV感染对3D4/2细胞活性的影响,将PRV分别以MOI=0.000111、0.00111、0.0111、0.111、1.11、11.1、111和1110接入3D4/2细胞,接毒2 h后弃上清液,继续培养4、8、12、24、48 h后测定细胞内或上清液中炎性因子水平和相关酶活性。结果显示,PRV可感染3D4/2细胞并形成明显细胞病变;当MOI=11.1、111和1110时,PRV感染3D4/2细胞48 h后显著抑制了细胞活性(P<0.05);与空白对照组相比,不同剂量PRV感染3D4/2细胞于不同时间点能够显著或极显著升高IL-1β、IL-6、IL-8、IL-10、TNF-α、IFN-γ、MCP-1、NO和ROS炎性因子水平及COX-1、COX-2、XOD、MPO和iNOS酶活性(P<0.05或P<0.01),极显著下调细胞内GSH含量水平(P<0.01)。结果表明,PRV能够感染3D4/2细胞,并且改变了3D4/2细胞的氧化还原状态,诱导3D4/2细胞产生炎症反应。将PRV按照MOI=0.111体外感染3D4/2细胞8 h~48 h是建立PRV感染3D4/2细胞氧化应激及炎症模型的最佳条件。The objective of this study was to observe the effects of PRV infection on oxidative stress and inflammatory response of porcine immune cells by infecting porcine alveolar macrophages(3d4/2)with pseudorabies virus(PRV),so as to lay a foundation for further study on the pathogenesis of PRV infection and the development of prevention and control drugs.PK-15 cells were used to propagate PRV,and the TCID_(50) of PRV was measured.The pathological lesions of 3D4/2 cells infected with PRV were observed.The effect of PRV infection on the activity of 3D4/2 cells was determined.3D4/2 cells were infected with PRV at MOI=0.000111,0.00111,0.0111,0.111 and 1.11 respectively.After two hours of inoculation,the supernatant was discarded,and the cells were further cultured for 4 h,8 h,12 h,24 h and 48 h.The levels of inflammatory factors and the activities of related enzymes in the cells or supernatant were measured.The results showed that when MOI=11.1,111 and 1110,PRV significantly inhibited the cell activity at 48 hours post infection(P<0.05),PRV could infect 3D4/2 cells and form obvious cytopathy.Compared with the control group,PRV infection significantly increased the levels of IL-1β,IL-6,IL-8,IL-10,MCP-1,TNF-α,IFN-γ,NO and ROS,and the activities of COX-1,COX-2,XOD,iNOS and MPO(P<0.05 or P<0.01),decreased intracellular GSH level in 3D4/2 cells(P<0.01).The results suggested that PRV could infect 3D4/2 cells,change the redox state of 3D4/2 cells,and induce inflammatory reaction in 3D4/2 cells.The best condition to establish the oxidative stress and inflammatory model of 3D4/2 cells was that PRV infected 3D4/2 cells at MOI=0.111 for 8-48 hours in vitro.

关 键 词：猪伪狂犬病病毒 猪肺泡巨噬细胞 氧化应激 炎症模型 炎症因子 

分 类 号：S852.659.1[农业科学—基础兽医学] S858.28[农业科学—兽医学]