基于上皮-间质转化探讨哮喘平冲剂干预支气管哮喘气道重塑的作用机制  

Investigating the mechanism of Xiaochuanping Granule in intervening airway remodeling in bronchial asthma based on epithelial-mesenchymal transition

作　　者：史梦蓉 张念志[2] 张芮 周静 孙小强 张倩倩 褚文丰 金纯纯 SHI Meng-rong;ZHANG Nian-zhi;ZHANG Rui;ZHOU Jing;SUN Xiao-qiang;ZHANG Qian-qian;CHU Wen-feng;JIN Chun-chun(First Clinical Medical School of Anhui University of Chinese Medicine,Hefei 230022,China;First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学第一临床医学院,安徽合肥230022 [2]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《时珍国医国药》2025年第4期609-615,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82174312);国家中医药管理局高水平中医药重点学科建设项目(国中医药人教函[2023]85号);安徽省中医肺病临床医学研究中心项目(皖科社[2020]41号)。

摘　　要：目的探究哮喘平冲剂基于上皮-间质转化(EMT)抑制哮喘模型大鼠气道重塑的作用机制。方法将70只雄性SD大鼠随机分为空白对照组、模型组、哮喘平低、中、高剂量组(3.87、7.74、15.48g·kg^(-1))、苏黄组(0.43g·kg^(-1))、地塞米松组(0.125mg·kg^(-1)),每组各10只。除空白对照组外,其他组均用卵清蛋白(OVA)注射致敏和雾化激发复制哮喘大鼠模型。各组大鼠连续21d灌胃相应药物,并观察其一般情况。苏木素-伊红(HE)染色和马松(Masson)染色观察各组大鼠肺组织及气道病理改变和胶原沉积情况;酶联免疫吸附测定法(ELISA)检测各组大鼠血清白介素17(IL-17)、白介素22(IL-22)、免疫球蛋白E(IgE)水平;实时荧光定量聚合酶链式反应(RT-qPCR)检测各组大鼠肺组织转化生长因子β1(TGF-β1)、Smad3、Smad7的mRNA表达情况;蛋白质免疫印迹(Western blot)检测各组大鼠肺组织中TGF-β1、Smad3、Smad7、α-平滑肌肌动蛋白(α-SMA)、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)表达水平。结果与空白对照组相比,大鼠模型组HE、Masson染色显示支气管及肺泡壁结构紊乱,上皮细胞变形、坏死,周围大量炎性细胞浸润,气道胶原纤维沉积明显增多,管壁增厚;血清IgE、IL-17、IL-22水平显著升高(P<0.01);肺组织TGF-β1mRNA、Smad3 mRNA及TGF-β1、Smad3、α-SMA及Vimentin表达显著上调(P<0.01),Smad7mRNA及Smad7、E-cadherin表达显著下调(P<0.01)。与模型组相比,哮喘平各剂量组、苏黄组、地塞米松组大鼠肺组织炎性细胞浸润及胶原纤维堆积程度减轻,气管壁厚度减少;血清IgE、IL-17、IL-22含量明显下降(P<0.05);肺组织TGF-β1mRNA、Smad3 mRNA及TGF-β1、Smad3、α-SMA及Vi⁃mentin表达显著下调(P<0.05),Smad7mRNA及Smad7、E-cadherin表达显著上调(P<0.05)。结论哮喘平冲剂可降低肺组织TGF-β1及Smad3表达水平,上调Smad7表达水平,抑制EMT,以改善哮喘气道重塑。Objective To investigate the mechanism by which Xiaochuanping Granule(XCPG)inhibits the airway remodeling in asthma model rats by regulating epithelial-mesenchymal transition(EMT).Methods Seventy male Sprague-Dawley(SD)rats were randomly divided into the blank control group,model group,XCPG low-dose,medium-dose,and high-dose groups(3.87g·kg^(-1)、7.74g·kg^(-1)、15.48g·kg^(-1)),Suhuang group(0.43g·kg^(-1)),with 10 rats in each group.Except for the blank control group,all the other groups were sensitized and then injected with ovalbumin(OVA)to create asthma in the rat models.Each rat group was given particular medications through a tube for 21 days in a row,and their general condition was observed.The pathological changes in the rat lung tis⁃sues and the deposition of airway collagen were observed using hematoxylin-eosin(HE)and Masson staining.Enzyme-linked immu⁃nosorbent assay(ELISA)was used to check the levels of IL-17,IL-22,and IgE in the blood of rats.Real-time fluorescent quantita⁃tive polymerase chain reaction(RT-qPCR)was used to check the mRNA expression levels of transforming growth factor Beta 1(TGF-β1),Smad3,and Smad7 in the lung tissues.Western blot(WB)was used to detect the expression levels of TGF-β1,Smad3,Smad7,alpha-smooth muscle actin(α-SMA),E-cadherin,and Vimentin in the lung tissues.Results Compared to the blank con⁃trol group,HE and Masson staining in the rat model group showed that the bronchial and alveolar wall structure was disordered,the epi⁃thelial cells were deformed and necrotic,a large number of inflammatory cells were infiltrated,the deposition of airway collagen fibers was significantly increased,and the tube wall was thickened;serum IgE,IL-17,and IL-22 levels were significantly elevated(P<0.01);TGF-β1 mRNA,Smad3 mRNA,and TGF-β1,Smad3,a-SMA,and wave protein expressions in the lung tissues were significantly upregulated(P<0.01),while Smad7 mRNA,Smad7,and E-cadherin expressions were significantly downregulated(P<0.01).Compared with the model group,in the XCPG groups

关 键 词：哮喘平冲剂 支气管哮喘 上皮-间质转化 气道重塑 TGF-β1/Smad3/Smad7 

分 类 号：R285.5[医药卫生—中药学] R256.12[医药卫生—中医学]