摘要
目的:采用网络药理学方法探索荆芥-防风药对抗过敏作用机制,并辅助体外细胞实验对相关靶点进行初步验证。方法:运用Cytoscape3.7.2软件,通过TCMSP、TTD、OMIM、Genecards和Uniprot数据库构建荆芥-防风药对活性成分-靶点交互网络;采用String数据库构建蛋白质相互作用(PPI)网络,运用DAVID在线数据库对交集靶点进行基因GO功能分析和KEGG通路富集分析。以抗-DNP-IgE致敏RBL-2H3细胞模型为载体,观察荆芥-防风药对乙酸乙酯萃取部位及分离物C、D对致敏细胞PI3K/AKT信号通路中相关蛋白表达量的影响。结果:共筛选出荆芥-防风药对潜在活性成分27个及相应靶点209个,过敏性疾病相关靶点2552个,交集靶点81个。GO富集分析结果显示,81个靶点主要涉及DNA模板转录、细胞增殖调控、细胞因子活性、酶结合等多种生物学过程。KEGG通路富集分析结果显示,PI3K/AKT信号通路、MAPK信号通路、Toll样受体信号通路等经典通路与过敏性疾病调控密切相关。细胞实验结果表明,荆防药对乙酸乙酯部位及其分离物C、D联用PI3K/AKT信号通路激活剂IGF-1,可下调致敏细胞内p-AKT蛋白表达量,而该通路抑制剂Wort则加强药物对PI3K、AKT、p-AKT蛋白表达的抑制作用。结论:荆芥-防风药对中槲皮素、木犀草素、汉黄芩素、β-谷甾醇、豆甾醇等活性成分可能是通过作用于AKT1、TNF、JUN、IL-4等81个靶点,及PI3K/AKT、MAPK、Toll样受体等主要信号通路来发挥抗过敏作用,且抑制PI3K/AKT信号通路的激活并展现了荆芥-防风药对抗过敏作用多成分-多靶点-多通路的作用特点。
Objective:To predict the anti-allergic mechanism of Schizonepeta tenuifolia Briq and Saposhnikovia divaricata(Trucz.)Schischk.(Jingjie-Fangfeng)by network pharmacology,and to verify preliminarily its related targets through vitro cell experiments.Methods:Cytoscape 3.7.2 software was used to construct an active ingredient-target interaction network through TCMSP,TTD,OMIM,Genecards and Uniprot databases.A String database was used to construct a protein interaction(PPI)network,and the DAVID online database was used to perform GO and KEGG enrichment analysis on intersection targets.An anti-DNP-IgE-sensitized RBL-2H3 cell model was used as carrier to observe the effect of ethyl acetate fraction extracted from Jingjie-Fangfeng drug pair and its isolate C,D on the expression of PI3K/AKT signaling pathway-related proteins in combination with tool drugs.Results:A total of 27 potential active ingredients and corresponding 209 targets were screened,as well as 2552 allergic diseases related targets,and 81 targets for intersection.The GO enrichment analysis showed that 81 targets were mainly involved in various biological processes such as DNA template transcription,cell proliferation regulation,cytokine activity,and enzyme binding.KEGG pathway enrichment analysis showed that the PI3K/AKT signaling pathway,MAPK signaling pathway and Toll-like receptor signaling pathway and other classical pathways had significant effects on the regulation of allergic diseases.The results of cell verification experiments showed that ethyl acetate fraction extracted from Jingjie-Fangfeng drug pair and its isolate C,D combined with PI3K/AKT signaling pathway activator IGF-1 down-regulated the expression of p-AKT protein in cells.While combined with the PI3K/AKT signaling pathway inhibitor Wort,the inhibition effect of drug pair on the expressions of PI3K/AKT pathway related proteins PI3K,AKT,p-AKT was enhanced Conclusion Jingjie-Fangfang drug pair has the anti-allegic effect,the active ingredients such as quercetin,luteolin,baicalein,β-sitoster
作者
刘淇
纪雅菲
周洪莉
吕红君
袁岸
方洋
曾南
Liu Qi;Ji Yafei;Zhou Hongli;Lv Hongjun;Yuan an;Fang Yang;Zeng Nan(Pharmacy College of Chengdu University of Traditional Chinese Medicine,Sichuan Province,Chengdu 611137;The Second Affiliated Hospital of Army Military Medical University,Chongqing 400000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2020年第5期136-143,共8页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金(编号:81473399)。
关键词
荆芥-防风药对
网络药理学
实验验证
抗过敏
靶点
Schizonepeta tenuifolia Briq.-Saposhnikovia divaricata(Trucz.)Schischk.drug pair
network pharmacology
anti-allergy
target
