基于网络药理学和分子对接探讨马齿苋改善肝损伤的分子作用机制

Exploring the Molecular Mechanism of Purslane in Improving Liver Injury Based on Network Pharmacology and Molecular Docking

目的基于网络药理学及分子对接技术探讨马齿苋改善肝损伤的有效活性成分及分子机制。方法通过Traditional Chinese Medicine Systems Pharmacology(TCMSP)数据库检索马齿苋的活性化合物及相关靶点;GeneCards、On⁃line Mendelian Inheritance in Man(OMIM)数据库收集肝损伤的相关靶点;通过Venny 2.1.0网站筛选马齿苋与肝损伤的共表达靶点。运用Cytoscape 3.9.1软件和Search Tool for the Retrieval of Interacting Genes/Proteins数据库构建马齿苋活性成分与肝损伤共表达靶点的“药物-主要活性成分-共表达靶点-疾病”网络以及蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络图。通过the Database for Annotation,Visu⁃alization and Integrated Discovery数据库对共表达靶点进行基因本体论(gene ontology,GO)富集和京都基因与基因组百科全书(Kyoto encycloppedia genes and genomes,KEGG)通路注释分析,阐述马齿苋改善肝损伤的作用通路及分子作用机制。通过CB-Dock在线分子对接网站验证马齿苋有效活性成分与关键靶点之间的关系。结果从TCMSP数据库中筛选出10种马齿苋活性化合物及其对应的78个靶点,从GeneCards和OMIM数据库中筛选出1489个肝损伤的相关靶点,通过Venny 2.1.0网站找到54个马齿苋活性成分与肝损伤的共表达靶点。通过Cytoscape 3.9.1软件获得“药物-主要活性成分-共表达靶点-疾病”网络并找出马齿苋改善肝损伤的主要活性成分,在PPI网络图中得到关键靶点为TNF、IL6、IL1B、MMP9、CASP3、IL10、PPARG。GO富集分析结果显示马齿苋主要对转录、凋亡等生物过程起调节作用,通过KEGG通路分析筛选出与马齿苋改善肝损伤关系密切的通路为Non-alcoholic fatty liver disease通路,并且其重要的靶点有TGFB1,INSR,TNF,TNFRSF1A,IL1A,CASP7,IL6,MAPK8,CASP8,IL1B,CASP3,PPARG和PPARA。分子对接结果表明,马齿苋中主要活性成分β-胡萝卜素、槲皮素与核心靶点TNF、CASP3、PPARG稳定结合。结论马齿苋中的β-胡萝卜素、槲皮素通过与TNF、CASP3、PPARG结合调节Non-alcoholic fatty liver disease等信号通路发挥改善肝损伤的作用。

Objective To explore the effective active ingredients and molecular mechanism of Portulaca oleracea in improving liver in⁃jury by network pharmacology and molecular docking technology.Methods The active compounds and related targets of Portulaca olera⁃cea were retrieved from the traditional Chinese medicine systems pharmacology(TCMSP)database.The related targets of liver injury were collected from GeneCards and Online Mendelian Inheritance in Man(OMIM)databases.The coexpression targets of Portulaca ol⁃eracea and liver injury were screened through Venny 2.1.0 website.Cytoscape 3.9.1 software and search tool for the retrieval of acting genes/proteins database were used to construct a“drug-main active ingredient-coexpression target-disease”network,and a proteinprotein interaction(PPI)network diagram of Portulaca oleracea active ingredients and liver injury coexpression targets.The Gene Ontol⁃ogy(GO)enrichment analysis and the Kyoto Encyclopedia Genes and Genomes(KEGG)pathway annotation analysis on the coexpres⁃sion targets were carried out through the Database for Annotation,Visualization and Integrated Discovery to elaborate the functional path⁃way and molecular mechanism of Portulaca oleracea in improving liver injury.The relationship between the effective active ingredients of Portulaca oleracea and their key targets was verified through CB-Dock online molecular docking website.Results Ten active com⁃pounds in Portulaca oleracea and their corresponding 78 targets were screened out from TCMSP database,1489 targets related to liver in⁃jury were screened out from GeneCards and OMIM databases,and 54 coexpression targets of Portulaca oleracea active ingredients and liver injury were found through Venny 2.1.0 website.The network of“drugs-main active ingredient-coexpression target-disease”was constructed through Cytoscape 3.9.1 software,and the main active ingredients of Portulaca oleracea to improve liver injury were found.In the PPI network diagram,the key targets were TNF,IL6,IL1B,MMP9,CASP3,IL10 and PPARG.The GO enrichment analysis showed that Portulaca oleracea mainly regulated biological processes such as transcription and apoptosis.Non-alcoholic fatty liver disease path⁃way,the pathway closely related to the improvement of Portulaca oleracea on liver injury,was screened out by KEGG pathway analysis,and the important targets in it were TGFB1,INSR,TNF,TNFRSF1A,IL1A,CASP7,IL6,MAPK8,CASP8,IL1B,CASP3,PPARG and PPARA.Molecular docking results showed that beta carotene and quercetin,the main active ingredients in Portulaca oleracea,were strongly bound to the core targets TNF,CASP3 and PPARG.Conclusion Beta carotene and quercetin in Portulaca oleracea may play a role in improving liver injury by binding to TNF,CASP3 and PPARG to regulate non alcoholic fatty liver disease signaling pathway.