摘要
目的:构建艾蒿花粉诱导小鼠过敏性哮喘模型并探讨其内在型,为后续精准治疗提供方法.方法:采用艾蒿花粉粗提物(MPE)腹腔致敏和滴鼻激发的方式构建BALB/c小鼠过敏性哮喘模型.将BALB/c小鼠随机分为PBS致敏PBS激发组(P-P)、MPE致敏PBS激发组(M-P)、MPE致敏MPE激发模型组(M-M).最后1次滴鼻激发24 h后,测定小鼠气道反应性;收集支气管肺泡灌洗液(BALF)进行细胞计数,流式细胞术对BALF各类炎症细胞进行分类统计;HE染色、PAS染色、Masson染色、α-SMA免疫组化染色观察肺组织病理学改变;ELISA检测肺组织和血清IFN-γ、IL-4、IL-5、IL-13和IL-17A,以及血清总IgE、MPE特异性IgE、IgG1和IgG2a水平.结果:MPE诱导小鼠呈现哮喘典型病理学特征,包括气道反应性升高、气道周围大量炎症细胞浸润、杯状上皮细胞化生增多、气道平滑肌增厚和气道周围纤维蛋白沉积等,BALF细胞总数由对照组的<1×105个/ml增加到模型组的>5×105个/ml,肺部炎症细胞浸润以嗜酸性粒细胞为主,且肺组织IL-4、IL-13和IL-17A、血清IL-5、IL-13水平均升高,血清总IgE水平及MPE特异性IgE、IgG1和IgG2a水平均显著升高.结论:MPE致敏激发可成功诱导小鼠以嗜酸性粒细胞浸润为主的过敏性哮喘,并伴有2型和17型炎症因子分泌增加,IgE、IgG1和IgG2a等抗体水平升高.
Abstract
Objective:To construct a mouse asthma model induced by mugwort pollen and to explore endotyping,providing methods for subsequent precision treatment.Methods:BALB/c mice were intraperitoneally injected with mugwort pollen extract(MPE)to sensitize,following MPE intranasal challenge to construct MPE allergic asthma murine model.Mice were randomly divided into PBS sensitization and PBS challenge(P-P),MPE sensitization and PBS challenge(M-P),MPE sensitization and MPE challenge model(M-M)groups.24 h after final challenge,mice were performed to examine airway responsiveness;bronchoalveolar lavage fluid(BALF)was harvested for cell counting and statistical classification of inflammatory cells through flow cytometry analysis.Pulmonary slides were collected for pathological examination,including HE,PAS,Masson and α-SMA immunohistochemical staining.ELISA was used to detect levels of IFN-γ,IL-4,IL-5,IL-13,IL-17A in lung tissue and serum,as well as serum total IgE and MPE-specific IgE,IgG1,IgG2a levels.Results:Pathological examination showed higher airway reactivity,more inflammatory cells infiltration around airway,obvious goblet metaplasia,thickening of airway smooth muscles and dramatical fibrin deposition around airway in model group.Total cell numbers of BALF were increased from<1×105 cells/ml in P-P group to>5×105 cells/ml in model group,in which eosinophils were predominant cellular type,levels of IL-4,IL-13,IL-17A in lung and IL-5,IL-13 levels in serum were significantly increased,as well as significant increasing levels of total IgE and MPE-specific IgE,IgG1,IgG2a.Conclusion:MPE-sensitized and challenged mice induces typical eosinophilic asthma featured with elevated eosinophils,as well as secretion of inflammatory factors of type 2 and type 17,IgE,IgG1 and IgG2a subtypes soars to high levels.
国家科技期刊平台
NSTL
维普
万方数据