三种证型对哮喘小鼠特异性/非特异性免疫功能及气道炎症的相关性研究
Correlation study of three types of syndromes on specific/nonspecific immune function and airway inflammation in asthmatic mice
投稿时间:2021-01-09  修订日期:2021-02-03
DOI:
中文关键词: 支气管哮喘  脾气虚证  肺气虚证  肺脾气虚证  免疫功能  气道炎症  气道阻力
英文关键词: Bronchial asthma  Spleen Qi Deficiency Syndrome  Lung Qi Deficiency Syndrome  Lung Spleen Qi Deficiency Syndrome  immune function  airway inflammation  airway resistance
基金项目:中南民族大学中央高校基本科研业务费专项资金项目(CZY20049,CZY19031)
作者单位E-mail
孙雅妮 中南民族大学 1135985443@qq.com 
陈诗 中南民族大学  
唐寒 中南民族大学  
葛月宾 中南民族大学 duckygreen@163.com 
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中文摘要:
      摘要 :目的:采用病证结合方法,构建三种证候哮喘小鼠模型,研究三种证型对哮喘小鼠特异性/非特异性免疫功能及气道炎症的影响及关联性研究。方法:健康的BALB/c实验小鼠,随机分为5组:对照组、哮喘组、脾气虚哮喘组、肺气虚哮喘组、肺脾气虚哮喘组。造模成功后,测定小鼠免疫器官指数;ConA,LPS诱导的T、B淋巴细胞增殖反应;流式细胞仪分析T细胞亚群CD3+、CD4+、CD8+的占比;支气管肺泡灌洗液(BALF)中白细胞分类计数;采用小鼠肺功能仪测定小鼠气道阻力(Rrs);苏木精-伊红(H&E)染色观察肺组织病理变化;酶联免疫吸附法(ELISA)测定小鼠BALF白介素-4(IL-4)、白介素-5(IL-5)、白介素-13(IL-13)以及血清中OVA特异性性免疫球蛋白E(OVA-IgE)、免疫球蛋白E(IgE)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)的含量。结果:与空白对照组相比,脾气虚哮喘组和肺脾气虚哮喘组免疫器官指数下降(P<0.05),三种证型T、B淋巴细胞增殖减少(P<0.05),T细胞亚型CD3+、CD4+占比减少(P<0.05),气道阻力增加(P<0.05) ,BALF中嗜酸性粒细胞、中性粒细胞、巨噬细胞增加(P<0.05)及IL-4、IL-5、IL-13的含量增加(P<0.05),血清中IgE含量增加(P<0.05)以及体液免疫指标IgA、IgG、IgM含量减少(P<0.05);模型组之间相比,肺脾气虚哮喘组免疫器官指数低于单纯哮喘组和肺气虚组以及T淋巴细胞增殖低于单纯哮喘组(P<0.05),肺气虚哮喘组T淋巴细胞增殖无显著性差异,三种证候哮喘模型T淋巴细胞亚型CD3+、CD4+占比低于单纯哮喘模型组(P<0.05),肺脾气虚哮喘组CD3+占比低于另外两组证候哮喘模型(P<0.05),肺脾气虚哮喘组嗜酸性粒细胞含量高于其他所有模型组(P<0.05)及气道阻力高于哮喘组和脾气虚哮喘组(P<0.05),肺脾气虚型哮喘组较其他三组模型血清中OVA-IgE浓度升高(P<0.05),血清中IgM水平显著低于肺气虚哮喘模型组(P<0.01)。结论:三种证型对哮喘小鼠特异性/非特异性免疫功能及气道炎症有影响且具有一定关联性。
英文摘要:
      Abstract:Objective: The combination of disease and syndrome was used to construct three syndromes of asthma mouse models to study the effects of the three syndromes on specific/non-specific immune function and airway inflammation in asthmatic mice and their correlation .Methods: Healthy BALB/c experimental mice were randomly divided into 5 groups: control group, asthma group, spleen qi deficiency asthma group, lung qi deficiency asthma group, lung spleen qi deficiency asthma group. After successful modeling, the mouse immune organ index was determined; ConA, LPS-induced proliferation of T and B lymphocytes; flow cytometry analysis of the proportions of T cell subsets CD3+, CD4+, CD8+; bronchoalveolar lavage fluid (BALF) ) Classification and count of white blood cells; Mouse lung function tester was used to measure mouse airway resistance (Rrs); Hematoxylin-Eosin (H&E) staining to observe lung tissue pathological changes; Enzyme-linked immunosorbent assay (ELISA) to measure mouse BALF Interleukin-4 (IL-4), Interleukin-5 (IL-5), Interleukin-13 (IL-13) and OVA-specific immunoglobulin E (OVA-IgE) and immunoglobulin E (IgE) in serum , Immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM) content . Results: Compared with the blank control group, the immune organ index of the spleen-qi deficiency asthma group and the lung-spleen-qi deficiency asthma group decreased (P<0.05), the proliferation of T and B lymphocytes of the three syndrome types decreased (P<0.05), and the T cell subtypes CD3+, The proportion of CD4+ decreased (P<0.05), airway resistance increased (P<0.05), eosinophils, neutrophils, and macrophages in BALF increased (P<0.05) and IL-4, IL-5, The content of IL-13 increased (P<0.05), the content of IgE in the serum increased (P<0.05), and the content of humoral immune indexes IgA, IgG, IgM decreased (P<0.05); compared with the model groups, lung-spleen-qi deficiency asthma The immune organ index of the group was lower than that of the simple asthma group and the lung-qi deficiency group and the proliferation of T lymphocytes was lower than that of the simple asthma group (P<0.05). There was no significant difference in the proliferation of T lymphocytes in the asthma group with lung-qi deficiency. The proportion of CD3+ and CD4+ of cell subtypes was lower than that of the simple asthma model group (P<0.05). The proportion of CD3+ in the asthma group with lung-spleen-qi deficiency was lower than the other two groups of syndrome asthma models (P<0.05). The level of granulocytes was higher than that of all other model groups (P<0.05) and the airway resistance was higher than that of the asthma group and spleen-qi deficiency asthma group (P<0.05). The lung-spleen-qi-deficiency asthma group was compared with the other three models in serum OVA-IgE concentration Increased (P<0.05), and the serum IgM level was significantly lower than that in the lung-qi deficiency asthma model group (P<0.01). conclusion: The three types of syndromes have effects on specific/non-specific immune function and airway inflammation in asthmatic mice and have certain correlations.
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