骨形态构建蛋白2型受体下调单核细胞趋化蛋白1/CC类趋化因子受体2通路对哮喘小鼠气道炎症和Th1/Th2平衡的影响  

Effect of BMPR2 on airway inflammation and the Th1/Th2 balance in asthmatic mice through downregulation of the MCP1/CCR2 pathway

作　　者：乔廉洁 敖铁 武俊慧 何世伟 蔡培 张振斌 韩新鹏 QIAO Lianjie;AO Tie;WU Junhui;HE Shiwei;CAI Pei;ZHANG Zhenbin;HAN Xinpeng(Department of Respiratory and Critical Care Medicine,Xi'an International Medical Center Hospital,Xi'an,Shaanxi 710021,China)

机构地区：[1]西安国际医学中心医院呼吸与危重症医学科,陕西西安710021

出　　处：《安徽医药》2024年第6期1124-1130,I0002,共8页Anhui Medical and Pharmaceutical Journal

基　　金：陕西省自然科学基础研究计划项目(2022JM-490)。

摘　　要：目的探究骨形态构建蛋白2型受体(BMPR2)对小鼠哮喘模型气道炎症和Th1/Th2平衡的影响及其机制。方法于2021年9月至2022年11月选用24只C57BL/6小鼠尾静脉注射BMPR2腺病毒,随后卵清蛋白(OVA)致敏和激发建立哮喘小鼠模型;采用随机数字表法分为对照组(生理盐水替代OVA造模)、OVA模型组(OVA诱导小鼠哮喘模型)、OVA+载体(Vector)组(空载慢病毒处理的OVA哮喘模型小鼠)、OVA+BMPR2组(BMPR2过表达慢病毒处理的OVA哮喘模型小鼠),每组6只。收集支气管肺泡灌洗液(BALF)和肺组织。免疫组织化学检测肺组织BMPR2表达水平;苏木精-伊红染色观察肺组织病理学改变;瑞氏染色后计数BALF中各类炎症细胞数目;酶联免疫吸附测定(ELISA)试剂盒检测BALF中白细胞介素(IL)-4,IL-5和IL-13炎症因子水平;蛋白质印迹法检测肺组织和气道上皮细胞16HBE中BMPR2、单核细胞趋化蛋白-1(MCP1)及其受体CC类趋化因子受体2(CCR2)蛋白表达水平。免疫共沉淀实验检测BMPR2与MCP1相互作用。结果与对照组相比,OVA模型组肺组织BMPR2(0.36±0.05比1.04±0.04)显著降低(P<0.01);小鼠肺泡破坏程度严重,肺组织有大量的淋巴细胞浸润;BALF中炎症细胞总数[(93.25±9.32)×10^(4)个/毫升比(4.79±0.41)×10^(4)个/毫升,P<0.001]、嗜酸性粒细胞、中性粒细胞和淋巴细胞均升高;Th1相关炎症因子γ干扰素(IFN-γ)水平降低,Th2相关炎症因子IL-4,IL-5和IL-13水平升高。过表达BMPR2可降低肺泡破坏程度和淋巴细胞浸润程度。与OVA+Vector组相比,OVA+BMPR2组BALF中炎症细胞总数、嗜酸性粒细胞、中性粒细胞和淋巴细胞均降低;ELISA结果表明,OVA+BMPR2组BALF中IFN-γ水平升高,IL-4,IL-5和IL-13水平降低,提示过表达BMPR2可上调Th1百分比,下调Th2细胞百分比。进一步研究表明,BMPR2过表达显著下调了MCP1及其受体CCR2的表达水平,且BMPR2与MCP1存在相互作用。结论BMPR2可通过下调MCP1/CCR2通路降低哮喘小鼠气�Objective To investigate the effects and underlying mechanisms of bone morphogenetic protein receptor 2(BMPR2)on airway inflammation and the Th1/Th2 balance in a mouse model of asthma.Methods From September 2021 to November 2022,24 C57BL/6 mice were selected and injected with BMPR2 adenovirus via the tail vein,followed by ovalbumin(OVA)sensitization and elicitation to establish an asthma mouse model;the mice were divided into a control group(saline alternative to OVA for modeling),an OVA model group(OVA-induced asthma model in mice),an OVA+Vector group(OVA asthma model mice treated with empty lentivirus),and an OVA+BMPR2 group(OVA asthma model mice treated with BMPR2-overexpressing lentivirus),with 6 mice in each group.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected.The expression level of BMPR2 in lung tissue was detected by immunohistochemistry;pathological changes in lung tissue were observed by hematoxylin-eosin staining;the number of various types of inflammatory cells in BALF was counted after Riehl's staining;the levels of interleukin(IL)-4,IL-5,and IL-13 inflammatory factors in BALF were detected by and enzyme-linked immunosorbent assay(ELISA)kit;and the levels of BMPR2,MCP1,and its receptor CClike chemokine receptor 2(CCR2)were detected by Western blotting in lung tissue and airway epithelial cells 16HBE.A coimmunoprecipitation assay was used to detect the interaction between BMPR2 and MCP1.Results Compared with those in the control group,BMPR2(0.36±0.05 vs.1.04±0.04)expression was significantly lower in the lung tissues of the OVA model group(P<0.01);alveolar de-struction was severe in the mice,and there was a large amount of lymphocyte infiltration in the lung tissues;the total number of inflam-matory cells in the BALF[(93.25±9.32)×10^(4)cells/mL vs.(4.79±0.41)×10^(4)cells/mL,P<0.001];eosinophils,neutrophils and lympho-cytes were elevated;the level of Th1-associated inflammatory factor interferon gamma(IFN-γ)was decreased;the levels of the Th2-as-sociated inflammatory factors IL

关 键 词：哮喘 骨形态构建蛋白2型受体 单核细胞趋化蛋白-1 气道炎症 辅助性T细胞1 辅助性T细胞2 小鼠 近交C57BL 

分 类 号：R562.25[医药卫生—呼吸系统] R-332[医药卫生—内科学]