LY294002干预对哮喘小鼠M2巨噬细胞极化及气道炎症的机制研究  

Mechanism of LY294002 intervention on M2 macrophage polarization and airway inflammation in asthmatic mice

作　　者：李丽蔚 陈慧雯 胡芳茴 梁伯灯 杨霞[1] Li Li-wei;Chen Hui-wen;Hu Fang-hui;Liang Bo-deng;Yang Xia(Department of General Medicine,The First Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi 530021,China)

机构地区：[1]广西医科大学第一附属医院全科医学科,广西南宁530021

出　　处：《中国现代医学杂志》2024年第24期29-35,共7页China Journal of Modern Medicine

基　　金：广西研究生教育创新计划项目(No:JGY2023080)。

摘　　要：目的探讨LY294002干预对哮喘小鼠M2巨噬细胞极化及气道炎症的机制研究。方法将24只雄性BALB/C小鼠随机分为空白对照组、哮喘模型组、LY294002组、地塞米松组,每组6只。采用腹腔注射卵清蛋白致敏及雾化吸入激发法复制小鼠哮喘模型,连续7 d腹腔注射给药后,观察小鼠肺组织中气道炎症、杯状细胞增生情况,酶联免疫吸附试验检测血清免疫球蛋白E(IgE)、白细胞介素-4(IL-4)、IL-25及支气管肺泡灌洗液中炎症细胞分类计数,实时荧光聚合酶链反应检测肺组织炎症因子IL-4、IL-13、IL-25R及M2标志物ARG-1、Ym-1 mRNA表达,Western blotting检测肺组织PI3K、p-PI3K、Akt、p-Akt及M2标志物CD206蛋白表达。结果哮喘模型组小鼠的支气管肺泡灌洗液中细胞总数、嗜酸性粒细胞、中性粒细胞、淋巴细胞均较空白对照组高(P<0.05),巨噬细胞较空白对照组低(P<0.05);LY294002组、地塞米松组细胞总数、嗜酸性粒细胞、中性粒细胞、淋巴细胞比例均较哮喘模型组低(P<0.05),巨噬细胞较哮喘模型组高(P<0.05)。哮喘模型组血清IgE、IL-4和IL-25水平均较空白对照组高(P<0.05),LY294002组、地塞米松组血清IgE、IL-4和IL-25水平均较哮喘模型组低(P<0.05)。哮喘模型组ARG-1、YM-1、IL-4、IL-13、IL-25R mRNA相对表达量均较空白对照组高(P<0.05),LY294002组、地塞米松组ARG-1、YM-1、IL-4、IL-13、IL-25R mRNA相对表达量均较哮喘模型组低(P<0.05)。哮喘模型组p-PI3K/PI3K、p-Akt/Akt、CD206蛋白相对表达量均较空白对照组高(P<0.05),LY294002组、地塞米松组p-PI3K/PI3K、p-Akt/Akt、CD206蛋白相对表达量均较哮喘模型组低(P<0.05)。结论LY294002可显著减低哮喘小鼠肺部组织的M2极化,减轻哮喘小鼠的气道炎症作用,其作用机制可能与PI3K/Akt信号通路有关。Objective To explore the mechanism of LY294002 intervention on M2 macrophage polarization and airway inflammation in asthmatic mice.Methods The 24 male BALB/c mice were randomly divided into the blank control group,the asthma model group,the LY294002 group,and the dexamethasone group,with 6 mice in each group.The asthma mouse model was established by sensitization through intraperitoneal injection of ovalbumin and provocation via aerosol inhalation.After 7 days of continuous intraperitoneal drug administration,the airway inflammation and goblet cell hyperplasia in the lung tissues of mice were observed.Serum immunoglobulin E(IgE),interleukin-4(IL-4),and IL-25 were detected by ELISA,and inflammatory cell counts in bronchoalveolar lavage fluid were determined.Quantitative real-time polymerase chain reaction was used to detect the mRNA expressions of inflammatory factors IL-4,IL-13,and IL-25R,as well as M2 markers ARG-1 and Ym-1 in lung tissues.Western blotting was performed to detect the protein expressions of PI3K,p-PI3K,Akt,p-Akt,and M2 marker CD206 in lung tissues.Results Compared with the blank control group,the asthma model group had a higher total number of cells and greater percentages of eosinophils,neutrophils and lymphocytes(P<0.05),but a lower percentage of macrophages in bronchoalveolar lavage fluid(P<0.05).The total number of cells and the percentages of eosinophils,neutrophils and lymphocytes were lower(P<0.05),but the percentage of macrophage was higher in the LY294002 group and the dexamethasone group compared with the asthma model group(P<0.05).The serum levels of IgE,IL-4 and IL-25 in the asthma model group were higher than those in the blank control group(P<0.05),while they were lower in the LY294002 group and the dexamethasone group than in the asthma model group(P<0.05).The relative mRNA expressions of ARG-1,YM-1,IL-4,IL-13 and IL-25R in the asthma model group were higher than those in the blank control group(P<0.05),while they were lower in the LY294002 group and the dexamethasone group than i

关 键 词：哮喘 LY294002 M2巨噬细胞 PI3K/AKT信号通路 

分 类 号：R562.25[医药卫生—呼吸系统]