副交感神经对支气管哮喘模型Th17型免疫反应的作用及其机制  

Effect and mechanism of parasympathetic nerve on Th17-type immune responses in bronchial asthma model

作　　者：王爱利 李琳[2] 万军[1] WANG Ai-li;LI Lin;WAN Jun(Department of Respiratory and Critical Diseases,Wuhan No.1 Hospital,Wuhan 430022,China;Department of General Medicine,Wuhan No.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院呼吸与危重症医学科,武汉430022 [2]武汉市第一医院综合医疗科,武汉430022

出　　处：《现代免疫学》2023年第6期473-478,共6页Current Immunology

基　　金：武汉市医学科研项目(WX20D25)。

摘　　要：为探讨副交感神经对卵清蛋白(ovalbumin,OVA)诱导哮喘小鼠气道炎症和Th17型免疫反应的影响和可能机制,将BALB/c小鼠随机分为正常对照组、哮喘组、GTS-21[α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor,α7nAChR)激动剂]干预组和α-银环蛇毒素(α-bungarotoxin,α-BGT)(α7nAChR拮抗剂)干预组。除对照组外,其余各组采用OVA构建小鼠哮喘模型,每次激发前GTS-21干预组经静脉注射GTS-21,α-BGT干预组皮下注射α-BGT。ELISA检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中TGF-β1和IL-6的浓度;H-E染色观察肺组织病理学变化;RT-PCR检测肺组织中c-Rel和IL-17的mRNA表达水平;Western blotting检测肺组织中c-Rel、IL-17和RORγt蛋白的表达水平;FACS检测各组小鼠脾脏单细胞悬液中Th17占CD4^(+)T细胞的百分比。结果显示,OVA刺激后肺组织中有大量炎症细胞的浸润;与哮喘组比较,GTS-21干预能显著降低BALF中TGF-β1和IL-6的水平(均P<0.01),下调肺组织中RORγt蛋白、c-Rel和IL-17的mRNA及蛋白表达水平(均P<0.05),逆转OVA诱导的CD4^(+)T细胞向Th17分化(P<0.01);α-BGT干预组TGF-β1浓度显著升高(P<0.05),IL-6的浓度无明显变化(P>0.05),RORγt蛋白、c-Rel和IL-17的mRNA及蛋白表达水平显著升高(均P<0.01),Th17占CD4^(+)T细胞的百分比显著升高(P<0.01)。由此,副交感神经能抑制炎症反应,调控哮喘模型中Th17型细胞免疫反应,为探索一种新的哮喘治疗策略提供了方向。To explore the effect and mechanism of parasympathetic nerve on ovalbumin(OVA)-induced bronchial inflammation and Th17 function in asthmatic mice,the BALB/c mice were randomly divided into the normal control group,asthma group,GTS-21(α7 nicotinic acetylcholine receptor[α7nAChR]agonist)intervention group andα-bungarotoxin(α-BGT,α7nAChR antagonist)intervention group.OVA was used to establish the asthma model in all mice except those in the control group.Before each stimulation,GTS-21 orα-BGT was intravenously or cutaneously injected into mice of the GTS-21 intervention group orα-BGT intervention group,respectively.The concentrations of TGF-β1 and IL-6 were detected by ELISA in the supernatant of bronchoalveolar lavage fluid(BALF).H-E staining was used to observe the pathological changes in lung tissues.The expression levels of c-Rel and IL-17 mRNA in lung tissues were detected by RT-PCR.The expression levels of c-Rel,IL-17,and RORγt proteins in lung tissues were detected by Western blotting.The ratio of Th17 to CD4^(+)T cells in the spleen monocytes suspension was detected by FACS.The results showed that massive inflammatory cells infiltrated the lung tissues after OVA stimulation.GTS-21 intervention significantly decreased TGF-β1 and IL-6 levels in BALF(both P<0.01),down-regulated the level of RORγt protein as well as the mRNA and protein levels of c-Rel and IL-17(all P<0.05).GTS-21 treatment reversed the OVA-induced differentiation of CD4^(+)T cells to Th17 type(P<0.01).On the other hand,TGF-β1 concentration in theα-BGT group was significantly increased(P<0.05)while IL-6 concentration was not significantly changed(P>0.05),RORγt protein,c-Rel and IL-17 mRNA and protein expression levels were significantly increased(all P<0.01),and the percentage of Th17 in CD4^(+)T cells was significantly increased(P<0.01).In conclusion,the parasympathetic nerve suppresses the inflammatory response and regulates the cellular immune response type Th17 in the asthma model,providing a good direction for exploring new th

关 键 词：副交感神经 哮喘 Α7烟碱型乙酰胆碱受体 辅助性T细胞17 

分 类 号：R562.2[医药卫生—呼吸系统]