cGAS蛋白促进PCV-2茎环结构诱导cGAS-STING信号通路天然免疫反应研究  

Research on cGAS promoted PCV-2 stem-loop structure to induce the innate immune response in the cGAS-STING signaling pathway

作　　者：苏冰 邓治邦[1] SU Bing;DENG Zhibang(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学动物医学院,长沙410128

出　　处：《黑龙江畜牧兽医》2024年第13期59-65,71,共8页Heilongjiang Animal Science And veterinary Medicine

摘　　要：为了探索猪圆环病毒2型(PCV-2)DNA茎环结构是否可以激活环鸟苷酸腺苷酸合成酶(cGAS),以及是否诱导cGAS-STING信号通路天然免疫反应,试验将含有PCV-2茎环结构的单链DNA(SL-DNA)转染至PK15细胞中,通过Western-blot检测cGAS蛋白是否在PK15细胞中表达;将SL-DNA转染至过表达及敲除cGAS蛋白的PK15细胞中,通过实时荧光定量PCR检测细胞中cGAS、干扰素刺激基因(STING)、TANK结合激酶1(TBK1)、干扰素调节因子3(IRF3)、干扰素-β(IFN-β)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)基因的表达水平。结果表明:SL-DNA转染后,PK15细胞中cGAS蛋白表达量上升;过表达cGAS蛋白的PK15细胞与PK15细胞相比,SL-DNA转染后cGAS、STING、TBK1、IRF3、IFN-β、IL-1β、IL-6、TNF-α基因的相对表达量均极显著升高(P<0.01);敲除cGAS蛋白的PK15细胞与PK15细胞相比,SL-DNA转染后cGAS、STING、TBK1、IRF3、IFN-β、IL-1β、IL-6、TNF-α基因的相对表达量均极显著降低(P<0.01)。说明cGAS蛋白可以识别PCV-2的茎环结构,并且促进其诱导的cGAS-STING信号通路天然免疫反应。In order to explore whether Porcine circovirus type 2(PCV-2)DNA stem-loop structure could activate cyclic guanosine adenylate synthase(cGAS)and whether it could induce a natural immune response to the cGAS-STING signaling pathway,in the test,single-stranded DNA(SL-DNA)containing PCV-2 stem-loop structure was transfected into PK15 cells,and whether cGAS was expressed in PK15 cells was detected by Western-blot.SL-DNA was transfected into cGAS-overexpressing and cGAS-knockout PK15 cells,and the gene expression levels of cGAS,interferon-stimulated gene(STING),TANK-binding kinase 1(TBK1),interferon-regulatory factor 3(IRF3),interferon-beta(IFN-β),interleukin-1beta(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected in the cells by realtime fluorescence quantitative PCR.The results showed that cGAS expression increased in PK15 cells after SL-DNA transfection.The relative gene expression of cGAS,STING,TBK1,IRF3,IFN-β,IL-1β,IL-6,and TNF-αwas extremely significantly elevated in cGAS-overexpressing PK15 cells compared with PK15 cells after SL-DNA transfection(P<0.01).The relative expression of extremely cGAS,STING,TBK1,IRF3,IFN-β,IL-1β,IL-6,and TNF-αgenes were all extremely significantly decreased(P<0.01)in cGAS-knockout PK15 cells compared with PK15 cells after SL-DNA transfection.The results suggested that cGAS recognized the stem-loop structure of PCV-2 and promoted its induced natural immune response by the cGAS-STING signaling pathway.

关 键 词：猪圆环病毒2型 茎环结构 环鸟苷酸腺苷酸合成酶 cGAS-STING信号通路 天然免疫反应 

分 类 号：S852.65[农业科学—基础兽医学]