基于网络药理学探讨哮喘宁颗粒治疗支气管哮喘的作用机制及PI3K/Akt信号通路验证  

Mechanism of Xiaochuanning Granules in Treatment of Bronchial Asthma via PI3K/Akt Signaling Pathway Based on Network Pharmacology and Experimental Verification

作　　者：黄帅阳 候丹 黄贵锐 吕明圣 弓雪峰 张诗瑜 张志杰 崔红生 HUANG Shuai-yang;HOU Dan;HUANG Gui-rui;LYU Ming-sheng;GONG Xue-feng;ZHANG Shi-yu;ZHANG Zhi-jie;CUI Hong-sheng(Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China;Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China)

机构地区：[1]北京中医药大学第三附属医院,北京100029 [2]北京中医药大学,北京100029 [3]首都医科大学附属北京朝阳医院,北京100020

出　　处：《中国实验方剂学杂志》2022年第9期150-157,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然基金面上项目(82074390)。

摘　　要：目的:基于网络药理学预测哮喘宁颗粒治疗支气管哮喘的潜在分子机制,并采用卵蛋白(OVA)致敏诱发的支气管哮喘大鼠模型对关键信号通路之一磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)进行实验验证。方法:利用传统中药系统药理学数据库和分析平台(TCMSP)和中药分子机制生物信息学分析网络(BATMAN-TCM)筛选哮喘宁颗粒的主要有效成分和作用靶点。通过人类基因数据库(GeneCards)、人类孟德尔遗传靶点数据库(OMIM)等5个疾病数据库获取支气管哮喘相关的疾病靶点。筛选并通过韦恩图获得两者的共同靶标,使用蛋白相互作用数据库(STRING)构建化合物-疾病的蛋白质-蛋白质相互作用网络(PPI),并用Cytoscape 3.8.0建立“哮喘宁关键活性成分-核心靶基因”网络。通过基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。制备OVA刺激诱导的支气管哮喘大鼠模型,通过苏木素-伊红(HE)染色观察支气管与肺组织炎症情况,蛋白免疫印迹法(Western blot)实验验证网络药理学富集分析结果。结果:该实验分别获取哮喘宁活性成分232个,相关靶点4687个,收集哮喘宁与支气管哮喘的共同靶点233个,包括嗜酸性粒细胞衍生神经毒素1(EDN1)、环磷腺苷效应元件结合蛋白1(CREB1)、细胞周期蛋白依赖性激酶抑制剂1A(CDKN1A)、表皮生长因子受体(EGFR)、丝裂原活化蛋白激酶14(MAPK14)、Akt1等。KEGG通路分析筛选了186条相关信号通路,显示PI3K/Akt信号通路可能在哮喘宁治疗支气管哮喘的过程中起关键作用。动物实验表明,与哮喘模型组比较,哮喘宁组可明显减轻大鼠气道炎症反应和平滑肌增生,并明显下调PI3K、Akt蛋白表达(P<0.05)。结论:哮喘宁治疗哮喘具有多成分、多靶点、多通路协同作用的特点,其中哮喘宁可能通过调控PI3K/Akt信号通路相关基因的表达,进一步发挥抗炎症的药理效应,为后续深入研究�Objective:To explore the underlying molecular mechanism of Xiaochuanning granules in the treatment of bronchial asthma based on the network pharmacology and experimental verification through the phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway on ovalbumin(OVA)sensitization-induced bronchial asthma model in rats.Method:The main active ingredients and targets of Xiaochuanning Granules were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM).The targets related to bronchial asthma were obtained from five disease databases such as GeneCards and Online Mendelian Inheritance in Man(OMIM).The common targets were screened out through the Venn diagram.STRING was used to construct the protein-protein interaction(PPI)network of"compound-disease",and Cytoscape 3.8.0 was used to establish a network of key active ingredients of Xiaochuanning granules and core target genes("ingredient-gene"network).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed through DAVID.The bronchial asthma model was induced by OVA stimulation in rats.Bronchial and lung tissue inflammation was observed by hematoxylin-eosin(HE)staining,and the enrichment analysis results of the network pharmacology were verified by Western blot.Result:In this experiment,232 active ingredients and 4687related targets of Xiaochuanning granules were screened out,and 233 common targets of Xiaochuanning granules and bronchial asthma were collected,including eosinophil-derived neurotoxin 1(EDN1),cyclic AMP response element-binding protein 1(CREB1),cyclin-dependent kinase inhibitor 1A(CDKN1A),epidermal growth factor receptor(EGFR),mitogen-activated protein kinase 14(MAPK14),and Akt1.KEGG pathway analysis revealed 186 related signaling pathways,indicating that the PI3K/Akt signaling pathway presumedly played a key role in the treatment of bronc

关 键 词：哮喘宁颗粒 支气管哮喘 网络药理学 动物验证 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]