鸟苷酸结合蛋白2b在牛分枝杆菌诱导巨噬细胞极化过程中的作用  

Role of Guanylate Binding Protein 2b during Macrophage Polarization Induced by Mycobacterium bovis

作　　者：于有利 王建东 郭亚男 张久盘 薛峰[2] 曹钰莹 YU Youli;WANG Jiandong;GUO Ya'nan;ZHANG Jiupan;XUE Feng;CAO Yuying(Institute of Animal Science,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002,China;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]宁夏农林科学院动物科学研究所,银川750002 [2]南京农业大学动物医学院,南京210095

出　　处：《畜牧兽医学报》2024年第6期2641-2651,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：宁夏回族自治区重大科技成果转化项目(2023CJE09028)。

摘　　要：旨在探究鸟苷酸结合蛋白2b (guanylate-binding protein 2b, GBP2b)对M.bovis感染的小鼠单核巨噬细胞系RAW264.7细胞M1、M2型极化的调控及其介导的信号通路。通过小干扰RNA下调RAW264.7巨噬细胞内GBP2b表达,分别利用qRT-PCR、Western blot和流式细胞术检测M1、M2型巨噬细胞表面标志物以及依赖MyD88信号通路关键蛋白表达情况;双萤光素酶报告基因检测GBP2b对NF-κB转录调控。结果表明GBP2b在M.bovis感染的巨噬细胞内高表达.下调RAW264.7巨噬细胞的GBP2b后再经M.bovis感染24 h, M1巨噬细胞表面标志物表达下调,而对M2巨噬细胞表面标志物表达无显著影响。下调GBP2b后抑制了NF-κB的转录启动,依赖MyD88信号通路相关蛋白随GBP2b的下调而下调。结果表明,GBP2b主要通过依赖MyD88信号通路促进M.bovis感染的RAW264.7巨噬细胞向M1表型极化和炎性反应,从而有助于巨噬细胞清除胞内M.bovis。This study aimed to investigate the effect of guanylate binding protein 2b(GBP2b)on M.bovis infection of RAW264.7 cells and its signal pathways in polarization of M1 and M2 during M.bovis infection.Downregulate the expression of GBP2b in RAW264.7 macrophages through small interfering RNA,and detect the expression of M1 and M2 macrophage surface markers and key proteins related to the MyD88 signaling pathway using qRT-PCR,Western blot,and flow cytometry,respectively.Dual luciferase reporter assay to examine the effect of GBP2b on NF-κB transcriptional regulation.The results showed that GBP2b was highly expressed in macrophages infected with M.bovis.After downregulating the expression of GBP2b in RAW264.7 macrophages and then being infected with M.bovis for 24 h,the expression of surface markers in M1 macrophages was downregulated,while there was no significant effect on the expression of surface markers in M2 macrophages.The transcription initiation of NF-κB is inhibited,and the downregulation of MyD88 signaling pathway related protein expression were due to the downregulation of GBP2b.The results indicate that GBP2b mainly promotes the polarization and inflammatory response of RAW264.7 macrophage infected with M.bovis towards the M1 phenotype by relying on the MyD88 signaling pathway,thereby helping macrophages clear intracellular M.bovis.

关 键 词：牛分枝杆菌 鸟苷酸结合蛋白2b 巨噬细胞 M1极化 M2极化 

分 类 号：S852.4[农业科学—基础兽医学]