射麻止喘液通过调控NF-κB/VEGFA信号通路抑制气道上皮细胞EMT改善哮喘气道重塑的机制研究  

tudy on mechanism of Shema Zhichuan Liquid for improving airway remodeling by inhibiting airway epithelial cell EMT in asthma through NF-κB/VEGFA signaling pathway

作　　者：张妙芬 王婷[1,2] 詹少锋[1] 黄慧婷[1] 江勇[3] 刘小虹 黄秀芳[1,2] ZHANG Miaofen;WANG Ting;ZHAN Shaofeng;HUANG Huiting;JIANG Yong;LIU Xiaohong;HUANG Xiufang(The First Affiliated Hospital,Guangzhou University of Chinese Medicine,Guangzhou 510000,China;Lingnan Medical Research Center of Guangzhou University of Chinese Medicine,Guangzhou 510000,China;Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine,Shenzhen 518072,China)

机构地区：[1]广州中医药大学第一附属医院,广州510000 [2]广州中医药大学岭南医学研究中心,广州510000 [3]深圳市中西医结合医院,深圳518072

出　　处：《中华中医药杂志》2024年第5期2569-2575,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.82204985);深圳市“医疗卫生三名工程”项目(No.SZZYSM202206013);广东省自然科学基金面上项目(No.2023A1515010807,No.2024A1515012183);广州市科技化项目(No.2023A04J1854);广东省教育厅项目(No.2022KQNCX013);广州中医药大学“双一流”与高水平大学协同创新团队项目(No.2021XK16);广东省重点科室(中西医协同科室)建设项目,深圳市中西医结合医院院内课题(No.YJ-2023-106)。

摘　　要：目的:探讨射麻止喘液对哮喘小鼠气道上皮细胞间充质转化(EMT)和气道重塑的影响及其潜在机制。方法:将30只SPF级雌性小鼠随机分为空白组,模型组,射麻止喘液低剂量、高剂量组,地塞米松组。屋尘螨滴鼻致敏建立哮喘小鼠模型。计算小鼠肺泡灌洗液(BALF)白细胞总数,HE、PAS、Masson染色法观察小鼠肺组织病理形态,ELISA法检测血清炎症因子和总IgE含量,RT-PCR检测气道重塑相关指标mRNA表达水平,免疫组化和荧光检测肺组织气道上皮细胞EMT标志物的表达情况,网络药理学预测射麻止喘液改善哮喘气道重塑的机制,Western Blot检测p-p65、p-IκBα、TNF-α、VEGFA关键蛋白的表达水平。结果:与模型组比较,3个给药组小鼠肺泡灌洗液中白细胞总数显著减少(P<0.01,P<0.05),血清IL-4、IL-5、总IgE含量显著降低(P<0.01,P<0.05),肺组织炎症细胞浸润、黏液分泌以及胶原沉积等病理情况显著改善(P<0.01),气道重塑相关指标Muc5ac、Fn1、α-SMA mRNA表达水平显著降低(P<0.01,P<0.05),肺组织中E-Cadherin、ZO-1蛋白表达水平升高,而N-Cadherin、α-SMA、p-p65、p-IκBα、TNF-α、VEGFA蛋白显著降低。结论:射麻止喘液可以通过抑制NF-κB/VEGFA信号通路改善气道上皮细胞EMT,进而改善哮喘气道重塑。Objective:To investigate the effects of Shema Zhichuan Liquid on epithelial-mesenchymal transition(EMT)and airway remodeling in asthmatic mice and its potential mechanisms.Methods:Thirty SPF-grade female mice were randomly divided into control group,model group,low-dose and high-dose Shema Zhichuan Liquid groups,and Dexamethasone group.House dust mite nasal drop sensitization was used to establish an asthma mouse model.The total number of white cells in bronchoalveolar lavage fluid(BALF)was counted by cell counter,the histopathological morphology of mouse lung was observed by HE,PAS and Masson staining,serum inflammatory factors and total IgE were detected by ELISA,and the mRNA expression level of airway remodeling-related indexes was detected by Real-Time PCR.Immunohistochemistry and fluorescence were used to detecte the expression of EMT markers in airway epithelial cells of lung tissues,and Network pharmacology was applied to predict the mechanism of Shema Zhichuan Liquid improving airway remodeling in asthma.Lastly,the expression level of key proteins of p-p65,p-IκBα,TNF-α,and VEGFA were detected by Western Blot.Results:Compared with the model group,the total number of white cell in the alveolar lavage fluid of mice in the low-dose and high-dose groups of Shema Zhichuan Liquid and the Dexamethasone group was significantly reduced(P<0.01,P<0.05),the levels of serum IL-4,IL-5,and total IgE were significantly reduced(P<0.01,P<0.05),and the pathologies such as the infiltration of inflammatory cells in the lung tissues,the secretion of mucus,and the deposition of collagen were significantly improved(P<0.01).The expression levels of Muc5ac,Fn1 andα-SMA m RNA,which were related the remodeling of the airways,were reduced(P<0.01,P<0.05),and the expression levels of E-Cadherin and ZO-1 proteins in lung tissues were elevated,whereas the proteins of N-Cadherin,α-SMA,p-p65,p-IκBα,TNF-α,and VEGFA were significantly reduced.Conclusion:Shema Zhichuan Liquid can inhibit airway epithelial cell EMT through NF-κB/VEGFA

关 键 词：哮喘 射麻止喘液 气道重塑 气道上皮细胞间充质转化 NF-κB/VEGFA信号通路 

分 类 号：R285.5[医药卫生—中药学]