IL-13对IRAK-M敲除哮喘小鼠TLR/IL-1R途径及巨噬细胞活化的影响  

Effects of IL-13 on TLR/IL-1R pathway and macrophage activation in IRAK-M knockout asthmatic mice

作　　者：胡朝阳 张小红 蒋天赐 刘风勋 李凤芝 刘倩倩 HU Chao-yang;ZHANG Xiao-hong;JIANG Tian-ci;LIU Feng-xun;LI Feng-zhi;LIU Qian-qian(Department of Respiratory and Critical Care Medicine,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000;Department of Nephrology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000,China)

机构地区：[1]郑州大学附属郑州中心医院呼吸与危重症医学科,河南郑州450000 [2]郑州大学附属郑州中心医院肾病科,河南郑州450000

出　　处：《热带医学杂志》2021年第11期1389-1395,共7页Journal of Tropical Medicine

基　　金：河南省高等学校重点科研项目(18A320056)。

摘　　要：目的探讨白介素(IL)-13对IL-1受体相关激酶M(IRAK-M)敲除哮喘小鼠巨噬细胞活化及Toll样受体/白介素1受体(TLR/IL-1R)途径的影响。方法实验分组:对照组、IRAK-M敲除组、哮喘组、IRAK-M敲除+哮喘组、哮喘+抗IL-13抗体组、IRAK-M敲除+哮喘+抗IL-13抗体组。收集肺泡灌洗液(BALF)并对细胞总数、嗜酸粒细胞、中性粒细胞、淋巴细胞、巨噬细胞计数;酶联免疫吸附(ELISA)检测BALF中IL-13、干扰素-γ(IFN-γ)、IL-4表达水平;实时荧光定量PCR(qRT-PCR)检测肺组织中甘露糖受体(MR)、精氨酸酶1(Arg1)、诱导性一氧化氮合酶(iNOS)mRNA水平;蛋白免疫印迹检测肺组织中IRAK-M、IL-13、TLR4、核转录因子kappaB(NF-κB)蛋白水平。结果与对照组相比,哮喘组各细胞数,BALF中IL-13、IL-4水平,肺组织中MR、Arg1、iNOS mRNA水平及IL-13、IRAK-M、TLR4、胞核NF-κB蛋白水平升高,差异有统计学意义(P<0.05);BALF中IFN-γ水平,肺组织中胞浆NF-κB蛋白水平降低,差异有统计学意义(P<0.05)。分别与IRAK-M敲除+哮喘组、哮喘+抗IL-13抗体组相比,IRAK-M敲除+哮喘+抗IL-13抗体组各细胞数,BALF中IL-13、IL-4水平,肺组织中MR、Arg1、iNOS mRNA水平及IL-13、IRAK-M、TLR4、胞核NF-κB蛋白水平降低,差异有统计学意义(P<0.05);BALF中IFN-γ水平,肺组织中胞浆NF-κB蛋白水平升高,差异有统计学意义(P<0.05)。结论抗IL-13抗体与IRAK-M敲除功能类似,均可缓解哮喘小鼠巨噬细胞活化状况。Objective To investigate the effects of interleukin(IL)-13 on macrophage activation and Toll like receptor/interleukin-1 receptor(TLR/IL-1 R)pathway in asthma mice with interleukin-1 receptor-associated kinase M(IRAK-M)knockout.Methods Bronchoalveolar lavage fluids(BALF)were collected from control group,IRAK-M knockout group,asthma group,IRAK-M knockout+asthma group,asthma+anti-IL-13 antibody group,IRAK-M knockout+asthma+anti-IL-13 antibody group;total number of cells,eosinophils,neutrophils,lymphocytes,macrophages cells were counted;the expression levels of IL-13,interferon-γ(IFN-γ)and IL-4 in BALF were detected by enzyme-linked immunosorbent assay(ELISA);the mRNA levels of mannose receptor(MR),arginase-1(Arg1)and inducible nitric oxide synthetase(iNOS)were determined by real-time fluorescence quantitative PCR(qRT-PCR);the protein levels of IRAK-M,IL-13,TLR4 and nuclear transcription factor kappa B(NF-κB)were detected by Western blot.Results Compared with the control group,the number of cells,the IL-13 and IL-4 levels in BALF,the MR,Arg1 and iNOS mRNA levels in lung tissue,the IL-13,IRAK-M,TLR4 and NF-κB protein levels in nucleus in asthma group were higher(P<0.05);and IFN-γ level in BALF and NF-κB protein level in cytoplasm of lung tissue were lower(P<0.05).Compared with IRAK-M knockout+asthma group and asthma+anti-IL-13 antibody group,the total number of cells,the IL-13 and IL-4 levels in BALF,the MR,Arg1 and iNOS mRNA levels in lung tissue,the IL-13,IRAK-M,TLR4 and NF-κB protein levels in nucleus in IRAK-M knockout+asthma+anti-IL-13 antibody group were lower(P<0.05);and IFN-γ level in BALF and NF-κB protein level in cytoplasm of lung tissue were higher(P<0.05).Conclusion The function of anti-IL-13 antibody was similar to that of IRAK-M knockout,it can alleviate the activation of macrophages in asthmatic mice.

关 键 词：白介素-13 白介素-1受体相关激酶M 哮喘 Toll样受体/白介素1受体途径 巨噬细胞活化 

分 类 号：R562.25[医药卫生—呼吸系统]