基于含药肝孵育液法的麻杏石甘汤抗气道上皮损伤的作用机制研究  

Mechanism of Action of Maxing Shigan Decoction Against Airway Epithelial Injury Based on Drug-Containing Liver Incubation Solution

作　　者：李文宏[1] 樊学程 雷婷 马嘉鑫 谢金根 陈林林 肖雄[1] 高文军[1] LI Wen-hong;FAN Xue-cheng;LEI Ting;MA Jia-xin;XIE Jin-gen;CHEN Lin-lin;XIAO Xiong;GAO Wen-jun(Collage of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学药学院,南昌330004

出　　处：《中国药学杂志》2022年第23期2013-2023,共11页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(82060790,81760785);江西省自然科学基金项目资助(20202ACBL206031);江西中医药大学校级研究生创新基金项目资助(JZYC21S31)。

摘　　要：目的结合体内外实验探讨含药肝孵育液法用于中药体外药理学研究的可行性及麻杏石甘汤抗气道上皮损伤的作用机制。方法SD雄性大鼠随机分为正常组、模型组、地塞米松组(dexamethasone,Dex)及麻杏石甘汤组(中药配比方A、B、C、D),麻杏石甘汤干预大鼠哮喘模型,记录引喘潜伏期,病理切片观察气管组织病理形态和气道上皮细胞超微结构变化,原位杂交和蛋白免疫印迹法(Western blot)分别检测表皮生长因子受体(EGFR)mRNA和蛋白表达;SD雄性大鼠离体气管随机分为正常组、模型组、布地奈德福莫特罗组(budesonide and formoterol fumarate powder for inhalation,BF)及麻杏石甘汤含药肝孵育液组(中药配比方A、B、C、D),麻杏石甘汤含药肝孵育液干预大鼠离体气管损伤模型,记录气管张力变化,酶联免疫吸附测定法(ELISA)检测气管EGFR、血管内皮生长因子(VEGF)和粒细胞-巨噬细胞集落刺激因子(GM-CSF)含量水平,原位杂交和免疫组化分别检测磷酸肌醇-3-激酶(PI3K)mRNA和蛋白表达;大鼠气道上皮细胞(RTE)随机分为正常组、模型组、地塞米松组及麻杏石甘汤含药肝孵育液组(中药配比方A、B、C、D),麻杏石甘汤含药肝孵育液干预RTE细胞损伤模型,流式细胞术检测RTE细胞损伤率,Western blot检测RTE细胞EGFR、PI3K蛋白表达。结果整体研究表明,与正常组比较,模型组气管组织EGFR mRNA和蛋白表达显著升高,气管组织结构形态和上皮细胞超微结构受损;与模型组比较,给药组均可延长大鼠引喘潜伏期,修复气管组织结构形态和上皮细胞超微结构损伤,并下调EGFR mRNA和蛋白表达,以A组药效最为显著。离体组织研究表明,与正常组比较,模型组气管EGFR、VEGF、GM-CSF含量水平显著升高,PI3KmRNA和蛋白表达显著升高;与模型组比较,给药组均可舒张气管环张力,降低EGFR、VEGF、GM-CSF含量水平,并下调PI3K mRNA和蛋白的表达,以A组药效最为显OBJECTIVE To explore the feasibility of using drug-containing liver incubation solution for in vitro pharmacology research of traditional Chinese medicine and the mechanism of action of Maxing Shigan decoction against airway epithelial injury combined with in vitro and in vivo experiments.METHODS SD male rats were randomly divided into normal group,model group,dexamethasone group(dexamethasone,Dex)and Maxing Shigan decoction group(herbal formula A,B,C,D).Maxing Shigan decoction was used to intervene in the rat model of asthma,the incubation period of asthma induction was recorded,the pathological changes morphology of tracheal histopathology and the ultrastructural of airway epithelial cells were observed by pathological section,in situ hybridization and Western blot were used to detect the mRNA and protein expression of epidermal growth factor receptor(EGFR).SD male rats with isolated trachea were randomly divided into the normal group,the model group,budesonide and formoterol fumarate powder for inhalation group(BF)and Maxing Shigan decoction-containing liver incubation solution group(herbal formula A,B,C,D).Maxing Shigan decoction-containing liver incubation solution intervened in the isolated tracheal injuryof rat model,the changes of tracheal tension was recorded,the levels of tracheal EGFR,vascular endothelial growth factor(VEGF)and granulocyte-macrophage colony-stimulating factor(GM-CSF)were detected by ELISA,and the mRNA and protein expressions of phosphoinositide-3-kinase(PI3 K)were detected by in situ hybridization and immunohistochemistry.Rat tracheal epithelial(RTE)cells were randomly divided into normal group,model group,dexamethasone group and Maxing Shigan decoction-containing liver incubation solution group(herbal formula A,B,C,D).Maxing Shigan decoction-containing liver incubation solution intervenes in RTE cells injury model,the damage rate of RTE cells was detected by flow cytometry,and the protein expressions of EGFR and PI3 K in RTE cells were detected by Western blot.RESULTS The overall expe

关 键 词：麻杏石甘汤 含药肝孵育液 支气管哮喘 气道上皮损伤 表皮生长因子受体/磷酸肌醇-3-激酶信号通路 气道重塑 

分 类 号：R966[医药卫生—药理学]