重组耻垢分枝杆菌减轻非嗜酸细胞性哮喘小鼠气道炎症的机制研究  

Mechanism of recombinant Mycobacterium smegmatis in alleviating airway inflammation in mice with non-eosinophilic asthma

作　　者：陈凌[1] 郭盛[2] 吴良霞[1] 张建华 Chen Ling;Guo Sheng;Wu Liangxia;Zhang Jianhua(Shanghai Jiao Tong University Affiliated Sixth People′s Hospital,Department of Pediatrics,Shanghai 200233,China;Children′s Hospital of Shanghai Jiao Tong University,Department of Endocrinology,Shanghai 200062,China;Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Department of Pediatrics,Shanghai 200092,China)

机构地区：[1]上海交通大学附属第六人民医院儿科,200233 [2]上海交通大学附属儿童医院内分泌科,200062 [3]上海交通大学医学院附属新华医院儿内科,200092

出　　处：《国际儿科学杂志》2019年第7期527-531,共5页International Journal of Pediatrics

基　　金：上海市卫生计生系统资助项目(2016ZB0102-03);上海市自然科学基金(19ZR1438400)

摘　　要：目的观察重组耻垢分枝杆菌(recombinant Mycobacterium smegmatis,rMS)-Ag85a-IL-17a鼻腔黏膜免疫对非嗜酸细胞性哮喘(non-eosinophilic asthma,NEA)小鼠气道炎症反应的影响,并探讨相关机制.方法SPF级DO11.10小鼠随机分为对照组(CON组)、哮喘组(OVA组)、重组耻垢分枝杆菌免疫组(rMS组).OVA混悬液、rMS菌液分别经鼻腔滴入建立NEA及rMS免疫干预动物模型.采用ELISA法测定小鼠血清中抗IL-17A自身抗体滴度及肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中IL-6和IL-17A浓度,流式细胞术(flow cytometry,FCM)检测BALF中性粒细胞比例;比色法检测肺组织匀浆髓过氧化物酶(myeloperoxidase,MPO)活力,实时定量PCR法测定肺组织CXCL2和CXCL5 mRNA表达.结果与正常对照组比较,哮喘组小鼠BALF中白细胞数量明显增加,以中性粒细胞为主,且IL-6和IL-17A表达显著升高,差异有统计学意义(P均<0.01);肺组织匀浆中MPO活力增强,CXCL2和CXCL5 mRNA表达上调,差异有统计学意义(P均<0.01).与哮喘组比较,rMS组小鼠血清中出现高滴度抗IL-17A自身抗体,BALF白细胞总数、中性粒细胞数量以及IL-6和IL-17A表达水平明显下降,肺组织MPO活力减低,CXCL2和CXCL5 mRNA表达显著下调,差异有统计学意义(P均<0.01).结论rMS经鼻腔黏膜免疫可在小鼠体内诱导产生抗IL-17A自身抗体,降低非嗜酸细胞性哮喘小鼠气道炎性细胞数量,抑制炎症因子表达.Objective To observe the effects of intranasal inoculation with recombinant Mycobacterium smegmatis(rMS)on the airway inflammation of non-eosinophilic asthmatic mouse and further investigate the relative mechanism.Methods DO11.10 T-cell receptor transgenic mice were divided randomly into three groups.Non-eosinophilic asthmatic model was established via OVA challenge,rMS were administrated into mice before challenge.Anti-IL-17A autoantibody in sera,IL-6 and IL-17A in BALF were measured by ELISA,the proportion of neutrophil in BALF was measured by FCM,MPO activity in lung tissue was detected by colorimetry,and the mRNA expression of CXCL2 and CXCL5 was measured by real-time PCR.Results Compared to control group,the number of neutrophils,IL-6 and IL-17A levels in BALF from asthmatic mice was significantly increased,meanwhile MPO activity and the expression of CXCL2,CXCL5 in lung tissue were both significantly upregulated.The results showed that high titer of autoantibody of IL-17A in sera of mice vaccined with recombinant Mycobacterium smegmatis was detected.Compared to asthma group,neutrophils and IL-6,IL-17A in BALF were significantly decreased,meanwhile MPO activity and CXCL2,CXL5 mRNA expression were significantly down-regulated.Conclusion Recombinant Mycobacterium smegmatis exhibits anti-inflammatory activity in murine neutrophilic asthma model and it may have protective effects on asthma.

关 键 词：支气管哮喘 IL-17A 自身抗体 气道炎症 

分 类 号：R56[医药卫生—呼吸系统]