BVDV感染对小鼠巨噬细胞IFN-β相关信号分子的影响  

Effects of BVDV infection on IFN-β-related signaling molecules in mouse macrophages

作　　者：段明媚 曹唱唱 赵心怡 陈斐 周彬 姜胜 邵春艳 周莹珊 董婉玉 杨杨 王晓杜 宋厚辉 宋泉江 DUAN Mingmei;CAO Changchang;ZHAO Xinyi;CHEN Fei;ZHOU Bin;JIANG Sheng;SHAO Chunyan;ZHOU Yingshan;DONG Wanyu;YANG Yang;WANG Xiaodu;SONG Houhui;SONG Quanjiang(Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Laboratory for Animal Health Inspection&Internet Technology/Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management/China-Australia Joint Laboratory for Animal Health Big Data Analytics,College of Animal Science and Technology/College of Veterinary Medicine,Zhejiang A&F University,Hangzhou 311300,China;Xinzhuang Animal Husbandry and Veterinary Station,Yantai,Shandong 265400,China)

机构地区：[1]浙江农林大学动物科技学院/动物医学学院、浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程实验室/动物医学与健康管理浙江省国际科技合作基地/中澳动物健康大数据分析联合实验室,浙江杭州311300 [2]山东省烟台市招远市辛庄畜牧兽医站,山东烟台265400

出　　处：《中国兽医学报》2023年第9期1828-1836,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金青年科学基金资助项目(31802258);浙江农林大学学校科研基金资助项目(2018FR015);浙江省自然科学基金资助项目(LY22C180003)。

摘　　要：为了探究致细胞病变型牛病毒性腹泻病毒(bovine viral diarrhealvirus,BVDV)对小鼠巨噬细胞IFN-β相关信号分子的影响,以0.1和3 MOI感染小鼠巨噬细胞RAW264.7,通过RT-qPCR、Western blot、ELISA等方法检测小鼠巨噬细胞中TLR37/TLR7、IRF3/IRF7、STAT1、IFN-β、ISG15/OAS1和USP18/SOCS1等因子的转录和表达水平差异,并对巨噬细胞中BVDV拷贝数进行检测。结果显示BVDV可以进入巨噬细胞细胞内,但病毒拷贝数不能随时间而增长。BVDV进入小鼠巨噬细胞先后上调TLR3和下调TLR7的转录水平,继而显著上调IRF3和IRF7转录水平,并呈现一定的剂量依赖性。STAT1和IFN-β的转录和表达也随BVDV感染而显著升高,而且IFN-β表达量随BVDV的病毒量和感染时间而增加。发挥抗病毒活性的ISG15和OAS1转录水平在BVDV感染前期出现显著升高,感染后期恢复正常,同时在感染后期干扰素负性调节因子USP18和SOCS1转录水平显著升高。所以致细胞病变型BVDV可以感染小鼠巨噬细胞,但不能复制,但可以通过TLR3/7-IRF3/7激活IFN相关信号通路,并接受负性调节因子的调控。To investigate the effect of cytopathic BVDV on IFN-βrelated signaling molecules of mouse macrophages,mouse macrophages RAW264.7 were infected with 0.1 MOI and 3 MOI BVDV.The transcription and expression levels of TLR37/TLR7,IRF3/IRF7,STAT1,IFN-β,ISG15/OAS1 and USP18/SOCS1 in mouse macrophages and BVDV copies in RAW264.7 were detected by RT-qPCR,Western blot,ELISA and other methods.The virus copy number did not increase with time in RAW264.7.When BVDV was introduced into mouse macrophages,the mRNA levels of TLR3and TLR7were up-regulated successively,and then the mRNA levels of IRF3and IRF7were significantly up-regulated in a dose-dependent manner.The mRNA and protein of STAT1and IFN-βwere also significantly increased with BVDV infection,and the expression of IFN-βincreased with the amount of BVDV virus and infection time.The mRNA levels of ISG15and OAS1,which exert antiviral activity,were significantly increased in the early stage of BVDV infection and recovered to normal in the later stage of infection.Meanwhile,the mRNA levels of interferon negative regulatory factors USP18and SOCS1 were significantly increased in the later stage of infection.The cytopathic BVDV could infect mouse macrophages,but could not replicate.The IFN related signaling pathway could be activated through TLR3/7-IRF3/7,and the activation response was regulated by inhibitors of IFN signaling.

关 键 词：牛病毒性腹泻病毒 小鼠 巨噬细胞 复制 干扰素 

分 类 号：S855.3[农业科学—临床兽医学]