基于cAMP信号通路探讨小青龙汤对CFTR离子通道调节哮喘黏液纤毛清除功能的作用机制  

Mechanism of Xiaoqinglong Decoction on Regulation of CFTR lon Channel on Mucociliary Clearance in Asthma Based on cAMP Signaling Pathway

作　　者：罗新越 王慧贤[2] 徐盼 黄东晖 LUO Xinyue;WANG Huixian;XU Pan;HUANG Donghui(The Second Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou Guangdong 510405,China;Guangdong Provincial Hospital of Chinese Medicine,Zhuhai,Zhuhai Guangdong 519015,China;Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine,Zhuhai Guangdong 519020,China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510405 [2]广东省中医院珠海医院,广东珠海519015 [3]珠海市中西医结合医院,广东珠海519020

出　　处：《新中医》2024年第14期14-19,共6页New Chinese Medicine

基　　金：广东省中医药局科研项目(20221361,20241290,20241285);北京晁恩祥中医药传承发展基金会2023年“迈向新天地”科研项目(2023CX03)。

摘　　要：目的:观察小青龙汤对哮喘寒饮蕴肺证小鼠环磷酸腺苷(cAMP)、囊性纤维化跨膜转导调节因子(CFTR)、黏蛋白5AC(MUC5AC)表达的影响,探讨小青龙汤基于cAMP通路对CFTR离子通道改善黏液纤毛清除功能的作用机制。方法:常规卵清白蛋白(OVA)加寒性刺激干预建立小鼠哮喘寒饮蕴肺证模型,于末次激发后取肺组织进行苏木素-伊红(HE)及阿尔辛蓝-过碘酸雪夫(AB-PAS)染色,收集支气管肺泡灌洗液及肺组织上清液,检测肺泡灌洗液中MUC5AC水平及肺组织上清液中cAMP、CFTR水平。结果:成功构建哮喘小鼠寒饮蕴肺证模型,小鼠肺泡灌洗液中MUC5AC表达上升,肺组织上清液中cAMP、CFTR表达均下降,肺组织病理切片见气道周围炎症细胞浸润,气道黏液分泌及杯状细胞增生。与模型组比较,地塞米松组、小青龙汤组肺泡灌洗液中MUC5AC表达均下降,小青龙汤组肺泡灌洗液中MUC5AC表达下降幅度小于地塞米松组,差异均有统计学意义(P<0.05)。与模型组比较,cAMP组、小青龙汤组肺组织上清液中cAMP、CFTR表达均上升,小青龙汤组肺组织上清液中cAMP、CFTR表达上升幅度均小于cAMP组,差异均有统计学意义(P<0.05)。病理切片见,小青龙汤组支气管周围基本无炎性细胞浸润,支气管管壁及肺泡保持较完整,平滑肌增生、支气管增厚与cAMP组、地塞米松组相比程度较轻,管腔内未见黏液栓及脱落的肺泡上皮。小青龙汤组支气管管壁清晰,杯状细胞增生较少,气道内黏液分泌少于cAMP组、地塞米松组。结论:小青龙汤可以减轻哮喘寒饮蕴肺证小鼠气道黏液高分泌及保障黏液纤毛清除功能运行,作用机制为通过cAMP通路提高CFTR离子通道。Objective:To explore the mechanism of Xiaoqinglong Decoction on improving mucociliary clearance function of CFTR ion channel based on cAMP pathway by observing the dynamic changes of cAMP,cystic fibrosis transmembrane conductance regulator(CFTR),and MUC5AC expression in asthma mice with syndrome of cold-rheum accumulation in the lung.Methods:Model mice with asthma with syndrome of cold-rheum accumulation in the lung were established by ovalbumin(OVA)plus cold stimulation.After the last stimulation,lung tissues were stained with hematoxylin-eosin(HE)and Alcian blue-periodic acid Schiff(AB-PAS).Bronchoalveolar lavage fluid and lung tissue supernatants were collected,and the levels of MUC5AC in bronchoalveolar lavage fluid and the levels of cAMP and CFTR in lung tissue supernatants were detected.Results:Model mice with asthma with syndrome of cold-rheum accumulation in the lung were successfully established.The expression of MUC5AC in bronchoalveolar lavage fluid was increased,and the expression of cAMP and CFTR in lung tissue supernatants was decreased.Histological sections showed inflammatory cells infiltration around the airways,mucus secretion and hyperplasia of goblet cells.Compared with that in the model group,the expression of MUC5AC in bronchoalveolar lavage fluid was decreased in the Dexamethasone group and the Xiaoqinglong Decoction group,and the decrease of expression of MUC5AC in bronchoalveolar lavage fluid in the Xiaoqinglong Decoction group was smaller than that in the Dexamethasone group,differences being significant(P<0.05).Compared with that in the model group,the expression of cAMP and CFTR in the lung tissue supernatants was increased in the cAMP group and the Xiaoqinglong Decoction group respectively,and the increase of expression of cAMP and CFTR in the lung tissue in the Xiaoqinglong Decoction group Was smaller than that in the cAMP group,differences being significant(P<0.05).According to histological sections,there was almost no inflammatory cell infiltration around the bronchi in the Xiaoqin

关 键 词：支气管哮喘 小鼠 寒饮蕴肺证 小青龙汤 环磷酸腺苷 囊性纤维化跨膜转导调节因子 黏蛋白5AC 

分 类 号：R259[医药卫生—中西医结合]