雾化吸入灭活草分枝杆菌对哮喘模型小鼠T-bet/GATA-3表达的影响  

Effect of inhalation inactived Mycobacterium phlei on T-bet/GATA-3 expression in asthmatic mice

作　　者：刘位位 孙起翔 张景鸿[3] 杨霞[3] 李超乾[1] LIU Wei-wei;SUN Qi-xiang;ZHANG Jing-hong;YANG Xia;LI Chao-qian(Dept of Respiratory Medicine;Dept of Emergency,the First Affiliated Hospital ofGuangxi Medical University,Nanning 530021,China;Dept of Respiratory Medicine,Jiangbin Hospital of Guangxi,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院呼吸内科,广西南宁530021 [2]广西壮族自治区江滨医院呼吸科,广西南宁530021 [3]广西医科大学第一附属医院急诊科,广西南宁530021

出　　处：《中国药理学通报》2018年第8期1099-1104,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81470230;81360007)

摘　　要：目的探讨雾化吸入灭活草分枝杆菌对哮喘小鼠Tbet/GATA-3表达的影响,了解草分枝杆菌的免疫调节机制。方法将24只♂Balb/c小鼠随机分为正常组、哮喘组、治疗组。模型组用鸡卵清蛋白(OVA)致敏激发制备哮喘小鼠模型,治疗组在OVA激发后雾化吸入灭活草分枝杆菌5 d,每日1次。各组小鼠处死后,取肺组织和支气管肺泡灌洗液(BALF),左肺组织病理切片行HE染色和过碘酸-雪夫染色(PAS染色),观察小鼠肺内炎症改变;ELISA法检测BALF中IFN-γ和IL-4水平;实时荧光定量PCR检测肺组织中Tbet、GATA-3 mRNA表达水平;流式细胞术检测肺单个核细胞Th1、Th2细胞占CD4+T细胞百分比。结果肺组织HE染色发现,与哮喘组相比,治疗组支气管周围炎症细胞浸润减少,PAS染色发现治疗组管腔中杯状细胞较哮喘组明显减少;哮喘组中IFN-γ、肺组织中T-bet mRNA表达水平和Th1细胞百分比与正常组相比明显降低(P<0.05),IL-4、GATA-3 mRNA表达水平和Th2细胞百分比较正常组明显升高(P<0.05);治疗组IFN-γ、肺组织中T-bet mRNA表达水平和Th1细胞百分比较哮喘组明显升高(P<0.05),IL-4、GATA-3mRNA表达水平和Th2细胞百分比较哮喘组明显降低(P<0.05)。相关性分析发现,小鼠肺组织中T-bet mRNA与Th1细胞百分比呈明显正相关(r=0.70,P<0.01),GATA-3 mRNA与Th2细胞百分比呈明显正相关(r=0.76,P<0.01)。结论雾化吸入灭活草分枝杆菌可以减轻哮喘小鼠气道炎症,通过调节转录因子水平,增加T-bet mRNA,降低GATA-3mRNA表达水平,进而抑制Th2细胞因子分泌。Aim To investigate the effect of inhalation of inactivated Mycobacterium phlei on the expression of T-bet/GATA-3 in asthmatic mice,and to understand the immunoregulatory mechanism of Mycobacterium phlei. Methods Twenty-four male Balb/c mice were randomly divided into normal group,asthma group and treatment group. Asthmatic model was established by ovalbumin（OVA）,and the treatment group was inactivated by Mycobacterium phlei after OVA challenge for five days,1 time a day. The lung tissues and bronchoalveolar lavage fluid（BALF） were harvested. Lung inflammation changes were observed by HE staining and periodic acid Schiff（PAS） staining. The levels of IFN-γ and interleukin（IL）-4 in BALF were measured by ELISA. The levels of T-bet,GATA-3 mRNA expressions in lung tissues were determined by real timePCR. The percentage of Th1 and Th2 cells in CD4＋T cells in lung mononuclear cells was detected by flow cytometry. Results Mycobacterium phlei treatment alleviated lung inflammation and attenuated mucus production. Compared with normal group, the asthma model group significantly decreased IFN-γ,T-bet mRNA levels and percentage of Th1 cells（P〈0. 05） and significantly increased IL-4,GATA-3 mRNA levels and percentage of Th2 cells（P〈0. 05）. Compared with asthma model group,treatment group significantly increased IFN-γ,T-bet mRNA levels and percentage of Th1 cells（P〈0. 05） and significantly decreased IL-4,GATA-3 mRNA levels and percentage of Th2 cells（P〈0. 05）. Correlation analysis showed that the levels of T-bet mRNA in lung tissues were positively correlated with the percentage of Th1 cells（r = 0. 7,P〈0. 01）,while the levels of GATA-3 mRNA were positively correlated with the percentage of Th2 cells（r = 0. 76,P〈0. 01）. Conclusions Inhalation of inactivated Mycobacterium phlei could alleviate airway inflammation in asthmatic mice. It can inhibit the secretion of Th2 cytokines by increasing T-bet mRNA expression,while inhibiting GATA-3 mRNA expression at transcriptional leve

关 键 词：哮喘 灭活草分枝杆菌 T-BET GATA-3 TH1 TH2 

分 类 号：R-332[医药卫生] R322.35