蓝莲清瘟败毒饮方药抗艾滋病病毒作用机制的网络药理学分析及实验验证  

Mechanism of Lanlian Recipe in the inhibition of HIV based on network pharmacology and experiment

作　　者：秦明珍 陈路[1] 刘钰[1] 蓝鸣生[1] 吕建楠 吴无畏[1] QIN Mingzhen;CHEN Lu;LIU Yu;LAN Mingsheng;LYU Jiannan;WU Wuwei(Guangxi Botanical Garden of Medicinal Plants,Nanning 530010,China;Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533099,China)

机构地区：[1]广西壮族自治区药用植物园,广西南宁530010 [2]右江民族医学院附属医院,广西百色533099

出　　处：《药学研究》2023年第12期969-977,共9页Journal of Pharmaceutical Research

基　　金：广西科学研究与技术开发计划项目(No.桂科AD17292003)。

摘　　要：目的以网络药理学方法分析蓝莲清瘟败毒饮方药(蓝莲方药)抗人类免疫缺陷病毒(HIV)的分子机制,并通过体外实验验证其抗病毒活性,为抗HIV的基础研究和新药研发提供参考。方法采用中药系统药理学数据库与分析平台(TCMSP)获取蓝莲方药各中药组分的化学成分及其靶点,进行药物动力学(ADME)筛选后将活性靶点导入Uniprot进行规范化;利用GeneCards、OMIM、TTD以及DrugBank数据库筛选HIV相关基因;将蓝莲方药调控HIV的关键基因输入Metascape平台分析其参与的主要生物学过程和信号通路;利用STRING平台以及Cytoscape 3.8.0软件构建蓝莲方药调控HIV的“成分-靶点-通路网络图”以及蛋白互作网络(PPI);选取主要成分与核心靶点通过AutoDock Vina 1.2.3软件进行分子对接验证;通过体外实验测定蓝莲方药对TZM-bl的细胞毒性,同时检测蓝莲方药抑制HIV假病毒感染细胞的活性;利用实时荧光定量PCR技术检测主要通路信号分子mRNA的表达水平。结果蓝莲方药抗HIV病毒的主要活性成分为槲皮素、木犀草素、山柰酚、黄芩素等,核心靶点有AKT1、TNF和CASP3等,关键生物学通路为PI3K-Akt信号通路、TNF信号通路和IL^(-1)7信号通路。分子对接结果显示主要成分与核心靶点具有良好的结合活性。细胞实验得出蓝莲方药和对照组唐草片的CC50分别为3.16、2.01 mg·mL^(-1);对HIV病毒的IC_(50)分别为17.10μg·mL^(-1),94.42μg·mL^(-1);蓝莲方药与唐草片相比具有更好的抗HIV作用(P<0.05)。实时荧光定量PCR结果显示蓝莲方药能够显著提高IL^(-1)7的mRNA表达水平。结论蓝莲方药可抑制HIV在TZM-bl细胞内的增殖,其中发挥主要作用的机制可能是对IL^(-1)7信号通路的调控。Objective The molecular mechanism of Lanlian Recipe in the inhibition of human immunodeficiency virus(HIV)was analyzed based on the network pharmacological method,providing a reference for the basic research and development of new drugs of anti-HIV.Methods The main chemical compositions of the components of Lanlian Recipe and the potential therapeutic targets were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).The potential targets were introduced into UniProt for standardization after ADME screening.The HIV related targets were screened form Genecards,OMIM,TTD and DrugBank databases.Metascape platform was applied to analyze the main biological processes and signal pathways of those HIV related targets.A"drug ingredients-targets-pathways"network and Protein-Protein Interaction Network(PPI)was constructed using Cytoscape 3.8.0 software and STRING database.The active ingredients of Lanlian Recipe were selected for molecular docking with the core targets through AutoDock Vina 1.2.3.The cytotoxicity to TZM-bl cell and the anti-HIV activity of Lanlian Recipe were determined in vitro.Results Our study showed that the main HIV-related active ingredients of Lanlian Recipe are quercetin,luteolin,kaempferol,baicalein and the core targets are AKT1,TNF,CASP3,etc.The main signaling pathways of Lanlian Recipe that regulating HIV are PI3K-AKT,TNF and IL^(-1)7 pathway.The molecular docking showed that a good binding activity between the active ingredients and core targets.The CC 50 of Lanlian Recipe and Tangcao Tablets for control that inhibiting HIV pseudo virus were 3.16 and 2.01 mg·mL^(-1),respectively,and the IC_(50) are 17.10 and 94.42μg·mL^(-1),respectively.These showed that Lanlian Recipe has a better anti-HIV activity than Tangcao Tablets(P<0.05).The result of quantitative real-time PCR revealed that Lanlian Recipe can significantly increase the expression of IL^(-1)7.Conclusion Lanlian Recipe can inhibit the proliferation of HIV in TZM-bl cells,and the main mecha

关 键 词：网络药理学 中药复方 蓝莲方药 人类免疫缺陷病毒 

分 类 号：R285[医药卫生—中药学]