基于网络药理学和分子对接技术研究芹黄素抗猪传染性胃肠炎病毒的作用  

Research on mechanism of apigenin against transmissible gastroenteritis virus in-fection based on network pharmacology and molecular docking

作　　者：张祎 汤宇心 史晨曦 胡慧[1,2] ZHANG Yi;TANG Yuxin;SHI Chenxi;HU Hui(College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;Ministry of Education Key Laboratory for Animal Pathogens and Biosafety,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046 [2]动物病原与生物安全教育部重点实验室,河南郑州450046

出　　处：《中国兽医学报》2025年第2期312-321,共10页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金重点资助项目(32130106)。

摘　　要：基于网络药理学和分子对接探究芹黄素抗猪传染性胃肠炎病毒(transmissible gastroenteritis virus, TGEV)感染的作用。综合运用Pharmmapper、Pubchem等多个数据库获取芹黄素的作用靶点,通过检索PubMed数据库获取了TGEV感染的相关作用靶点,利用Draw Venn Diagram分析得到芹黄素-TGEV感染的共同靶点,通过STRING数据库分析蛋白质-蛋白质相互作用关系(PPI)并获取关键的核心靶点,采用DAVID数据库对共同靶点进行GO功能和KEGG通路富集分析,最后,通过分子对接和体外细胞试验对分析结果进行验证。在筛选过程中,共获得了431个芹黄素的作用靶点,1 177个TGEV感染的相关靶点以及50个芹黄素-TGEV感染的共同靶点;GO富集分析结果显示,芹黄素主要调控细胞分化、细胞膜筏形成、凋亡过程及炎症反应的调节;KEGG富集分析获得了前15条在统计学上具有显著性差异的结果,涉及到PI3K-Akt和TNF信号通路等;分子对接的结果显示,芹黄素与基质金属蛋白酶3(MMP3)结合能最低(-9.5 kJ/mol),结合活性最好。在TGEV感染LLC-PK1细胞的体外模型上进一步评价,结果显示,与病毒单独感染组相比,不同浓度芹黄素处理后能够显著降低病毒的滴度、病毒的基因拷贝数(P<0.01),显著降低MMP3及其下游蛋白(IL-1β)的表达量,显著降低p65和IκBα的磷酸化水平。综上所述,芹黄素可以通过多靶点、多通路抑制TGEV感染,且可能是通过作用于MMP3及其上、下游蛋白调控NF-κB-MMP3-IL-1β信号通路发挥作用。The aim of this study is to explore the mechanism of apigenin against transmissible gas-troenteritis virus(TGEV)infection based on network pharmacology and molecular docking.The potential targets of apigenin were obtained from Pharmmapper,Pubchem and other databases.The PubMed database was searched to obtain the relevant targets of TGEV infection.The intersection targets of apigenin and TGEV infection were identified by Draw Venn Diagram online program.A"drug-disease-target"network was constructed using STRING database and Cytoscape 3.8.2 soft-ware.Protein-protein interaction relationships were obtained from the STRING database,and core targets were analyzed.The intersection targets were subjected to GO function and KEGG pathway enrichment analysis using the DAVID database.Finally,the analysis results were validated through molecular docking and in vitro cell experiments.The study identified 431 targets for apigenin,1177 targets for TGEV infection,and 50 intersection targets for apigenin and TGEV infection.GO enrichment analysis indicated that apigenin was mainly involved in regulating cell differentiation,cell membrane raft formation,apoptosis,and inflammatory responses.The top 15 statistically sig-nificant KEGG enrichment results mainly involve the PI3K-Akt signaling pathway and TNF signa-ling pathway.Docking analysis showed that apigenin had the strongest interaction with matrix metalloproteinase 3(MMP3)with an affinity of-9.5 kJ/mol and the binding activity of MMP3 was the best.The results of in vitro experiments demonstrated that treatment of different concen-trations of apigenin significantly reduced virus titers,virus genome copies,and the expression lev-els of MMP3 and its upstream and downstream proteins compared to the virus-infected group.Api-genin may exert its anti-TGEV effects through multiple targets and pathways,possibly by regula-ting the NF-κB-MMP3-IL-1βsignaling pathway.

关 键 词：网络药理学 芹黄素 猪传染性胃肠炎病毒 抗病毒感染 

分 类 号：S852.65[农业科学—基础兽医学]