绵羊肺炎支原体及其脂质相关膜蛋白诱导绵羊肺泡巨噬细胞IL-1β表达的分子机制  

Expression of IL-1β in sheep Alveolar macrophages by Mycoplasma ovipneumoniae and LAMPs-treated

作　　者：白帆[1,2] 吴金迪 王晓晖 吕天星[1] 王艳芳[1] 郝永清 BAI Fan;WU Jin-di;WANG Xiao-hui;LYU Tian-xing;WANG Yan-fang;HAO Yong-qing(Laboratory of Microbiology and Immunolgy,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Veterinary Research Institute,Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences,Hohhot 010031,China;Key Laboratory of Clinical Diagnosis and Treatment Technology for Animal Epidemic Disease,Laboratory of Veterinary Pharmacology,Ministry of Agriculture,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学兽医学院,微生物与免疫学实验室,内蒙古呼和浩特010018 [2]内蒙古农牧业科学院兽医所,内蒙古呼和浩特010031 [3]内蒙古农业大学兽医学院农业部兽医药理学实验室/动物疫病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2020年第9期1746-1753,共8页Chinese Journal of Veterinary Science

基　　金：内蒙古农业大学科技成果转化专项资金资助项目(YZGC2017027);内蒙古自治区科技计划资助项目(201802066)。

摘　　要：为证明TLR2是否参与绵羊肺炎支原体(Mo)或LAMPs刺激肺泡巨噬细胞时细胞因子的表达,使用Mo或LAMPs刺激绵羊肺泡巨噬细胞后,利用荧光定量PCR、流式细胞术、Western blot以及ELISA等方法分别对TLR2转录和翻译水平、MAPK信号通路激活情况和IL-1β分泌情况进行检测。同时,使用TLR2抑制抗体及MAPK抑制剂后,利用荧光定量PCR、Western blot以及ELISA检测MAPK信号通路激活情况和IL-1β分泌情况。结果显示,绵羊肺泡巨噬细胞受Mo或LAMPs刺激后,TLR2在基因和蛋白表达水平均有不同程度上升,MAPK信号通路中各信号蛋白均发生磷酸化反应,IL-1β在基因和蛋白表达水平均有不同程度上升。对TLR2及MAPK功能抑制后,MAPK信号通路的激活及IL-1β的表达均受到明显抑制。结果提示,Mo及其LAMPs可通过绵羊肺泡巨噬细胞的TLR2受体激活MAPK信号通路,继而引起细胞因子IL-1β的分泌增多。To determine whether TLR2 is involved in cytokine expression when Mycoplasma ovipneumoniae(Mo)or LAMPs stimulate sheep alveolar macrophages,after infection of alveolar macrophages with Mo or LAMPs,the expression of TLR2 at the transcription and translational levels,activation of MAPK signaling pathway and secretion of IL-1βwere detected by RT-qPCR,flow cytometry,Western blot and ELISA,respectively.Meanwhile,after functional inhibition with TLR2 and MAPK inhibitors,the activation of MAPK signaling pathway and the secretion of IL-1βwere detected by RT-qPCR,Western blot and ELISA.Results showed that after cell stimulation,the expression level of TLR2 in gene and protein increased to different degrees,phosphorylation reaction occurred in all signaling proteins in the MAPK signaling pathway,and the expression level of IL-1βalso increased to different degrees.After the inhibition of TLR2 or MAPK functions,the activation of MAPK signaling pathway and the expression of IL-1βwere significantly inhibited.The results suggest that the host cytokine production is activated in response to Mo or Mo-derived LAMPs through the MAPK signalling pathway via TLR2.

关 键 词：绵羊肺炎支原体 脂质相关膜蛋白 丝裂原活化蛋白激酶 TOLL样受体2 IL-1Β 

分 类 号：S852.62[农业科学—基础兽医学]