PRV感染小鼠三叉神经节细胞后通过RIG-Ⅰ信号通路调控Ⅰ型干扰素分泌  

Regulation of type Ⅰ interferon secretion via the RIG-Ⅰ signaling pathway after PRV infection of mouse trigeminal ganglion cells

作　　者：廖正波 汤德元[1] 曾智勇[1] 王彬[1] 黄涛[1] 陈旭 袁盛林 何松 周飘 毛茵茗 LIAO Zhengbo;TANG Deyuan;ZENG Zhiyong;WANG Bin;HUANG Tao;CHEN Xu;YUAN Shenglin;HE Song;ZHOU Piao;MAO Yinming(College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025

出　　处：《中国兽医学报》2025年第2期255-265,共11页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31860716);贵州省科技支撑计划资助项目(黔科合支撑[2021]一般162)。

摘　　要：为探究伪狂犬病病毒(pseudorabies virus, PRV)感染小鼠三叉神经节(TG)细胞后对其抗病毒免疫信号通路及Ⅰ型干扰素因子(IFN-Ⅰ)的影响。本试验使用1 MOI PRV接种感染TG原代细胞,使用106.29TCID50PRV滴鼻感染小鼠,使用实时荧光定量PCR(qPCR)、Western blot和ELISA等技术检测小鼠TG原代细胞不同攻毒时间点及体内攻毒组和对照组的基因转录、蛋白表达及IFN-α和IFN-β的分泌情况。结果显示,PRV感染小鼠TG原代细胞后引起RIG-I、MAVS和IRF3的基因及蛋白表达变化,诱导了体内外IRF3和IκBα的磷酸化。ELISA结果显示,PRV感染后可调控小鼠TG原代细胞及小鼠TG内IFN-α和IFN-β的分泌。使用siRNA干扰TG细胞内RIG-Ⅰ表达后Western blot检测RIG-Ⅰ信号通路相关蛋白表达结果及IFN-α和IFN-β的分泌情况。结果显示,siRIG-Ⅰ成功干扰TG细胞内RIG-Ⅰ蛋白表达,并引起下游MAVS和IRF3等蛋白表达发生变化,同时调控TG细胞内IFN-α和IFN-β的分泌。研究结果表明,PRV感染可诱导小鼠TG原代细胞及小鼠TG内RIG-Ⅰ的表达,并通过RIG-Ⅰ-MAVS-IRF3信号轴调控TG细胞内IFN-Ⅰ的抗病毒免疫反应,以及PRV会抑制TG内IRF3表达,并在感染后期显著上调IFN-β的表达,这可能是PRV在感染后期导致小鼠快速死亡的重要原因。本试验揭示了PRV感染小鼠TG细胞后通过RIG-Ⅰ-MAVS-IRF3信号通路调控IFN-Ⅰ的部分抗病毒免疫机制,以及发现IRF3蛋白在体内外不同的变化趋势,为后续深入研究奠定了基础。This study investigates the effects of pseudorabies virus(PRV)infection on the antiviral immune signaling pathways and typeⅠinterferon factors in mouse trigeminal ganglion(TG)cells.In this experiment,primary TG cells were infected with PRV at a multiplicity of infection(MOI)of 1,while mice were infected via a drop-nose method using 106,29 TCID50 of PRV.Real-time fluorescence quantitative PCR(qPCR),Western blot and ELISA were used to assess gene tran-scription,protein expression,and the secretion of IFN-αand IFN-β.The results indicated that PRV infection of mouse TG primary cells led to alterations in the gene and protein expression of RIG-Ⅰ,MAVS,and IRF3,as well as the phosphorylation of IRF3 and IKBαboth in vivo and ex vivo.ELISA results showed that PRV infection could regulate the secretion of IFN-αand IFN-βin mouse primary TG cells and mouse TGs.The results of RIG-Ⅰsignaling pathway-related proteins and the secretion of IFN-a and IFN-βwere analyzed using Western blot after using siRNA to interfere with RIG-Ⅰexpression in TG cells.The results showed that siRIG-Ⅰsuccessfully inter-fered with RIG-Ⅰprotein expression in TG cells and caused changes in the expression of down-stream proteins such as MAVS and IRF3,and also regulated the secretion of IFN-αand IFN-βin TG cells.Furthermore,the results indicated that PRV infection induced the expression of RIG-Ⅰin mouse TG progenitor cells,regulating the antiviral immune response of typeⅠinterferon factors in TG cells through the RIG-Ⅰ-MAVS-IRF3 signaling axis.Notably,PRV inhibited the expression of IRF3 in TG cells while significantly upregulating the expression of IFN-βduring the later stages of infection,which may be an important factor in the important reason for the rapid mortality ob-served in mice during the late stages of PRV infection.This experiment elucidates part of the anti-viral immune mechanism mediated by the RIG-Ⅰ-MAVS-IRF3 signaling pathway in regulating typeⅠinterferon factor after PRV infection of mouse TG cells,as well a

关 键 词：PRV RIG-Ⅰ IRF3 信号通路 Ⅰ型干扰素 小鼠 

分 类 号：S852.4[农业科学—基础兽医学]