基于网络药理学和实验验证分析止哮汤治疗哮喘的分子机制  

Molecular Mechanism of Zhixiao Decoction(止哮汤)in the Treatment of Asthma Based on Network Pharmacology and Experimental Verification

作　　者：张凯月 李春楠 张楠茜 高晓晨 程端端 申嘉明 王跃龙 吕经纬 孙佳明 ZHANG Kaiyue;LI Chunnan;ZHANG Nanxi;GAO Xiaochen;CHENG Duanduan;SHEN Jiaming;WANG Yuelong;LV Jingwei;SUN Jiaming(Jilin Ginseng Research Institute,Changchun University of Chinese Medicine,Changchun 130114)

机构地区：[1]长春中医药大学吉林省人参科学研究院,长春130113

出　　处：《中药药理与临床》2023年第12期46-53,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：吉林省科技发展计划项目(编号:YDZJ202201ZYTS177);吉林省中医药科技项目(编号:2022011);吉林省教育厅产业化项目(编号:JJKH20210992KJ)。

摘　　要：目的:通过网络药理学和实验验证对止哮汤治疗哮喘的主要活性成分及潜在作用机制进行探讨。方法:运用TCMSP数据库获取止哮汤各药味活性成分,通过Swiss Target Prediction和GeneCards、OMIM、TTD数据库结合,获得止哮汤治疗哮喘的共有靶点。运用String数据库构建蛋白互作的PPI网络图,运用CytoNCA插件提取网络中的核心效应靶点,运用Cytoscape 3.7.1软件构建“药物-成分-核心效应靶点”的网络图,Metascape网站对核心效应靶点进行GO与KEGG功能的富集分析。分子对接由Autodock Vina软件完成。通过RT-PCR在LPS诱导人支气管上皮细胞(16HBE)炎症模型中检测止哮汤活性部位对16HBE细胞相关蛋白的表达。结果:从数据库中筛选得到止哮汤187个活性成分及治疗哮喘的124个共有靶点。提取得到19个核心效应靶点,包含AKT1、EGFR、SRC、等。GO功能富集分析发现止哮汤对氧化应激,蛋白质结合等分子过程,膜微域、膜筏等细胞组成,激素受体结合等分子功能产生影响。KEGG通路富集分析得到55条通路,主要包括松弛素信号通路、内分泌抵抗、ErbB信号通路、等。分子对接结果显示关键靶点与活性成分异鼠李素、槲皮素、木犀草素均有较好的结合活性。体外细胞实验表明,止哮汤膜分离组分B能显著恢复16HBE细胞活力,RT-PCR试验结果表明,止哮汤膜分离组分B可以下调Akt1、Egfr、Src、Pi3k mRNA的表达,并降低细胞凋亡率。结论:本研究初步证明止哮汤可以通过下调关键蛋白的表达来抑制气道炎症,达到治疗哮喘的效果,为探索抗止哮汤治疗哮喘的机制提供依据。Objective:To explore the main active ingredients and potential mechanism of Zhixiao Decoction(止哮汤,ZXT)in treating asthma through network pharmacology and experimental verification.Methods:The TCMSP database was used to obtain the active ingredients of ZXT.Swiss Target Prediction,along with GeneCards,OMIM,and TTD databases,was employed to identify the common targets of ZXT in treating asthma.The String database was used to construct a protein-protein interaction(PPI)network,and the CytoNCA plugin was used to extract core effect targets from the network.Cytoscape 3.7.1 software was used to build a network diagram of the“drug-ingredient-core effect target”.The Metascape was used for GO and KEGG enrichment analyses of core effect targets.Molecular docking was performed using Autodock Vina software.The activity of the active ingredients of ZXT on mRNA related to bronchial epithelial cell(16HBE)cells was detected in an LPS-induced 16HBE inflammation model using RT-PCR.Results:A total of 187 active ingredients of ZXT and 124 common targets for treating asthma were screened from the database.Nineteen core effect targets were extracted,including AKT1,EGFR,and SRC.GO enrichment analysis revealed that ZXT influenced molecular processes(e.g.,oxidative stress and protein binding),cellular components(e.g.,membrane microdomains and lipid rafts),and molecular functions(e.g.,hormone receptor binding).KEGG pathway enrichment analysis identified 55 pathways,including the relaxation signaling pathway,endocrine resistance,and the ErbB signaling pathway.Molecular docking results showed that key targets and active ingredients such as isorhamnetin,quercetin,and luteolin had good binding activity.In vitro cell exper-iments demonstrated that membrane-separated component B of ZXT significantly restored the vitality of 16HBE cells.RT-PCR results indicated that membrane-separated component B of ZXT could inhibit the mRNA expression of Akt1,Egfr,Src,and Pi3k,and reduce cell apoptosis rate.Conclusion:This study reveals the scientific

关 键 词：止哮汤 网络药理学 分子对接 哮喘 作用机制 

分 类 号：R285[医药卫生—中药学]