甘露消毒丹调控PI3K/Akt信号通路干预甲型流感病毒感染的作用机制  

Mechanism of Ganlu Xiaodu Dan's intervention in influenza A virus infection by regulating PI3K/Akt signaling pathway

作　　者：黄廷轩 郭姗姗[2] 赵荣华[2] 张悦 吕思缘 于会勇 王成祥 HUANG Tingxuan;GUO Shanshan;ZHAO Ronghua;ZHANG Yue;LYU Siyuan;YU Huiyong;WANG Chengxiang(Department of Respiratory and Critical Care Medicine,Dongzhimen Hospital Xiamen Hospital,Beijing University of Chinese Medicine,Xiamen 361015,China;Biosecurity Laboratory,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700;Respiratory Department,the Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029)

机构地区：[1]北京中医药大学东直门医院厦门医院呼吸与危重症医学科,厦门361015 [2]中国中医科学院中药研究所生物安全实验室,北京100700 [3]北京中医药大学第三附属医院呼吸科,北京100029

出　　处：《北京中医药》2024年第8期871-879,共9页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82074389)。

摘　　要：目的 基于网络药理学探讨甘露消毒丹治疗甲型流感病毒感染的活性成分及作用机制,并用动物实验加以验证。方法 利用中药系统药理学数据库与分析平台(TCMSP)获取和筛选药物有效成分,利用TCMSP及Swiss Target Prediction数据库预测潜在作用靶点。利用比较毒物基因组学数据库(CTD)、疾病相关基因与突变位点数据库(DisGeNET)、GeneCards数据库、在线人类孟德尔遗传数据库(OMIM)获得甲型流感病毒感染的疾病靶点,将药物预测潜在作用靶点与疾病靶点取交集获得交集靶点。利用Cytoscape3.9. 1构建药物-成分-靶点网络并进行拓扑属性分析,预测中药主要活性成分。利用蛋白互作数据库(STRING)及Cytoscape3.9. 1绘制蛋白质-蛋白质相互作用(PPI)网络图,获取关键作用靶点。利用Metascape数据库进行基因本体(GO)功能以及京都基因和基因组百科全书(KEGG)通路富集分析。将C57BL/6J小鼠分为空白组、模型组、中药组及对照组,每组5只,甲型流感病毒PR8滴鼻感染小鼠构建感染模型,中药组予以甘露消毒丹灌胃,对照组予磷酸奥司他韦灌胃。观察肺指数、肺组织病理变化,实时荧光定量逆转录PCR (RTqPCR)法检测肺组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B (AKT)基因表达,双抗体夹心酶联免疫吸附法(ELISA)检测肺组织IL-6、IL-1β、TNF-α。结果 共筛选出甘露消毒丹125个化合物作为候选有效成分,获得286个潜在作用靶点,2 223个甲型流感病毒相关疾病靶点,61个交集靶点。药物-成分-靶点网络得到槲皮素、山奈酚、汉黄芩素、木犀草素、黄芩素、葛花苷元、金合欢素、异鼠李素、去氢二异丁香酚、柚皮素等10个关键成分。PPI网络得到TNF、IL-6、IL-1β、AKT1、TP53、JUN、HIF1A、PTGS2、MMP9、ESR1等10个关键靶点。GO功能富集分析共获得条目54条,其中生物过程20条,细胞组分14条,分子功能20条;KEGG通路富集分析共获得条目共20�Objective To investigate the active ingredients and mechanism of Ganlu Xiaodu Dan in the treatment of influenza A virus infection based on network pharmacology,and to verify it by animal experiments.Methods Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database were used to obtain and screen the active ingredients,and TCMSP and Swiss Target Prediction database were used to predict the potential targets.Comparative Toxicogenomics Database(CTD),Disease Gene Network(DisGeNET)database,GeneCards database and Online Mendelian Inheritance in Man(OMIM)database were used to obtain the disease targets of influenza A virus infection,and the potential targets of drug prediction were intersected with the disease targets to obtain the intersection targets.The drug-ingredient-target network was constructed by using Cytoscape3.9.1 and topological attribute analysis was performed to predict the main active ingredients of traditional Chinese medicine.Protein-protein interaction(PPI)network was mapped using Search Tool for the Retrieval of Interaction Gene/Proteins(STRING)database and Cytoscape3.9.1 to obtain key targets.Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)path enrichment were analyzed using Metascape database.C57BL/6J mice were divided into blank group,model group,traditional Chinese medicine group and oseltamivir group,with 5 mice in each group.Mice infected with influenza A virus PR8 nasal drops were constructed to establish the infection model.The traditional Chinese medicine group was given Ganlu Xiaodu Dan with gavage administration and the control group was given oseltamivir phosphate with gavage administration.V bThe lung index and lung tissue pathology were observed.The expression levels of Phosphatidylinositol 3-kinase(PI3K)and Protein kinase B(AKT)gene were detected by quantitative reverse transcription PCR(RT-qPCR),and the expression levels of IL-6,IL-1βand TNF-αwere detected by enzyme linked immunosorbent assay(ELISA).Results A total of 1

关 键 词：网络药理学 甲型流感病毒 靶点 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 甘露消毒丹 温病 

分 类 号：R285[医药卫生—中药学]