摘要
目的通过研究人抑制性细胞因子白细胞介素37b(interleukin 37b,IL-37b)转基因(transgenic,TG)小鼠(Il37b TG)对卵清蛋白(ovalbumin,OVA)诱导的哮喘气道高反应性、气道炎症和气道重塑等指标的变化,以及IL-37b在体外对小鼠和人肺结构细胞的作用,评估IL-37b在治疗哮喘方面的潜在意义。方法采用野生型和Il37b TG的雌性C57BL/6小鼠,建立周期为24 d的OVA诱导的哮喘小鼠模型,检测其肺力学参数、炎性细胞分类和Th2细胞因子表达以及肺组织病理学改变。体外采用原核细胞表达系统,获得大量纯化的重组人IL-37b。继而分别预处理小鼠肺成纤维细胞系(MLg)和人肺泡上皮细胞系(A549细胞),随后加以脂多糖(lipopolysaccharide,LPS)刺激,收集培养上清,采用双抗体夹心ELISA检测炎性细胞因子IL-6含量。结果与野生型OVA模型组相比,Il37b TG OVA组的气道高反应性降低,差异有统计学意义(P=0.0001,P<0.05);支气管肺泡灌洗液和肺组织中嗜酸性粒细胞数量降低,差异有统计学意义(分别为P=0.0001和P<0.0001,P<0.05);肺组织炎症细胞浸润明显减少,气道炎症、杯状细胞增生和黏液分泌均明显减轻,苏木精-伊红染色和过碘酸-希夫染色评分也有所降低,差异有统计学意义(分别为P=0.0001和P=0.0002,P<0.05),马松染色显示急性期胶原沉积的改变不明显;肺组织匀浆中Th2型细胞因子IL-5和IL-13的含量均有所降低。同时研究采用原核细胞表达系统得到的IL-37b预处理鼠肺成纤维细胞MLg和人肺上皮细胞A549,可以降低LPS所诱导IL-6的产生。结论IL-37b可能通过对相关靶细胞的负向调控进而发挥对哮喘气道的高反应性及气道炎症的抑制作用,为IL-37b作为一种新的治疗方法在临床治疗哮喘的潜在应用提供了一定的实验依据。
Objective We aimed to study the human inhibitory cytokine IL-37 b on the airways hyper-responsiveness and airway inflammation in an asthma model induced by ovalbumin(OVA)using IL-37 b transgenic mice(Il37b TG).It also attempted to investigate whether IL-37 b has inhibitory effects on structural cells of murine and human lungs,by in vitro cellular cultures in this study.The evaluation of the potential significance of IL-37 b in treatment of asthma was carried out by these experiments.Methods Using wild-type and Il37b TG female C57 BL/6 mice,with a 24-days circle,the asthma model was developed by OVA-induction,in which lung mechanical parameters,inflammatory cell classification in bronchoalveolar lavage fluid,Th2 cytokine expression,and histopathological changes of lung were examined.Prokaryotic cell expression system was developed in vitro to obtain purified recombinant human IL-37 b,which was used in pre-treatment of mouse lung fibroblast cell line(MLg)and human alveolar epithelial cell line(A549),respectively,and then stimulated with lipopolysaccharide(LPS).The culture supernatants were collected and the inflammatory cytokine IL-6 was tested by a double antibody sandwich ELISA.Results Compared with the wild-type OVA challenged group,the mice of the Il37b TG OVA group had significantly reduction in the airway hyper-responsiveness(P<0.0001,P<0.05),the number of eosinophils both in the bronchoalveolar lavage fluid(P=0.0001,P<0.05)and the lung tissues(P<0.0001,P<0.05),the inflammatory cells infiltrating into the lung tissues inflammation(P<0.0001,P<0.05)and mucus secretion(P=0.0002,P<0.05).Masson staining showed that the collagendeposition in the model on day 24 was not in obvious.The contents of Th2 type cytokines including IL-5 and IL-13 in lung tissue homogenate were also reduced in the mice of Il37b TG OVA group.In addition,the prokaryotic cell expression system used in this study was capable of obtaining a large quantity of highly purified human IL-37 b.The induction from IL-37 b pretreatment of MLg and A549 co
作者
秦啸峰
李艳
李陈铎
韦荣飞
李慎涛
袁慧慧
王晶晶
陈彦
刘杰
吕喆
孙英
徐大模
王炜
QIN Xiao-feng;LI Yan;LI Chen-duo;WEI Rong-fei;LI Shen-tao;YUAN Hui-hui;WANG Jing-jing;CHEN Yan;LIU Jie;LÜZhe;SUN Ying;XU Da-mo;WANG Wei(Department of Immunology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;不详)
出处
《微生物学免疫学进展》
2020年第5期6-15,共10页
Progress In Microbiology and Immunology
基金
国家自然科学基金项目(81700026、81971510)
