UPLC-Q-TOF/MS结合网络药理学和分子对接技术探讨山豆根抗肝癌的药效物质及作用机制

Pharmacodynamic material basis and mechanism of Sophorae Tonkinensis Radix et Rhizoma against liver cancer:an exploration based on UPLC-Q-TOF/MS,network pharmacology and molecular docking

关键词:  山豆根  肝癌  成分鉴定  网络药理学  分子对接

Key words:  Sophora tonkinensis  liver cancer  component identification  network pharmacology  molecular docking

摘要:

[目的]基于超高效液相色谱串联四级杆飞行时间质谱(UPLC-Q-TOF/MS)技术、网络药理学和分子对接技术探讨山豆根抗肝癌的药效物质基础和作用机制。[方法]采用UPLC-Q-TOF/MS技术鉴定对山豆根化学成分;运用PharmMapper、Swiss target prediction数据库预测山豆根成分靶点;通过GeneCards、DrugBank数据库收集肝癌相关靶点;运用Venny 2.1.0构建韦恩图,取化学成分和肝癌的交集靶点,并运用Cytoscape 3.7.2构建“化合物-靶点”网络,筛选山豆根抗肝癌的潜在成分;通过STRING数据库进行蛋白互相作用(PPI)网络分析,得到核心靶点;采用DAVID数据库对山豆根抗肝癌的核心靶点进行基因本体分析(GO)和京都基因和基因组百科全书(KEGG)富集通路分析,构建“化合物-靶点-通路”网络;山豆根抗肝癌的潜在活性成分与核心靶点蛋白的结合活性采用分子对接技术进行验证;通过细胞活性实验验证三叶豆紫檀苷、氧化苦参碱、苦参碱的抗肝癌作用。[结果] UPLC-Q-TOF/MS鉴定出山豆根中23个化学成分;获得山豆根成分和疾病的交集靶点33个。PPI分析、GO功能富集和KEGG通路富集分析发现,山豆根可能通过苦参碱、氧化苦参碱、6,8-二异戊烯基山柰酚、丁香脂素、Shandougenine A等活性成分作用于细胞周期素D1(CCND1)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、酪氨酸激酶受体2(ERBB2)、表皮生长因子受体(EGFR)、雌激素受体1(ESR1)等核心靶点,通过调节低氧诱导因子-1(HIF-1)信号通路、高级糖基化终末产物-受体(AGE-RAGE)信号通路、磷脂酰肌醇3激酶蛋白激酶B(PI3K/AKT)信号通路、酪氨酸激酶受体(ErbB)信号通路等多条通路发挥抗肝癌作用。分子对接验证得到山豆根潜在活性成分与核心靶点有较好结合力。细胞活性实验表明三叶豆紫檀苷、氧化苦参碱、苦参碱均有不同程度的抗肝癌作用。[结论]该研究初步表明山豆根通过多成分、多靶点、多途径发挥治疗肝癌的作用,为阐明山豆根抗肝癌的药效物质基础及作用机制研究提供了参考依据。

Abstract:

[Objective] To explore the pharmacodynamic material basis and mechanism of Sophorae Tonkinensis Radix et Rhizoma against liver cancer based on ultra-high pressure liquid chromatography tandem four-stage rod time-of-flight mass spectrometry(UPLC-QTOF-MS/MS) technique,network pharmacology and molecular docking. [Methods] UPLC-Q-TOF/MS technique was used to quickly identify the chemical components of Sophorae Tonkinensis. Pharm Mapper and Swiss Target Prediction were used to collect and predict component targets,followed by the search of liver cancer-related targets with GeneCards and DrugBank. The intersection targets of components of Sophorae Tonkinensis and liver cancer were obtained using Venny 2.1.0. Cytoscape 3.7.2 was used to construct the“chemical component-target-disease” network,to screen potential active components of Sophorae Tonkinensis against liver cancer.Protein-protein interaction(PPI) network was constructed using STRING to uncover the core targets of Sophorae Tonkinensis against liver cancer.,which were subjected to Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis based on DAVID database. and the “chemical component-target-pathway” network was constructed. The binding activity of the active components to the target proteins of Sophorae Tonkinensis against liver cancer and core target proteins was verified by molecular docking technology. Verify the anti hepatoma effect of Trifolirhizin,Oxymatrine and Matrine by cell activity test. [Results] UPLC-Q-TOF/MS revealed 23 ingredients of Sophorae Tonkinensis and a total of 33 intersection targets of Sophorae Tonkinensis components and the disease were obtained,PPI,GO,and KEGG analyses showed that Sophorae Tonkinensis presumedly acted on cyclin D1(CCND1),serine/threonine kinase 1(AKT1),receptor tyrosine kinase 2(ERBB2),epidermal growth factor receptor(EGFR),estrogen receptor 1(ESR1),and other targets through active components such as matrine,oxymatrine,6,8-diprenylkaempferol,syringaresinol and shandougenine A,and regulated hypoxia-inducible factor(HIF-1),receptor for advanced glucation end products(AGE-RAGE),phosphatidylinositol 3-kinase and protein kinase B(PI3K/AKT),receptor tyrosine kinase(ErbB),and other signaling pathways to play the role of anti hepatoma.Molecular docking verified a good binding ability of the potential active ingredients of Sophorae Tonkinensis to the core targets. Cell activity test showed that Trifolirhizin,Oxymatrine and Matrine had different anti hepatoma effects. [Conclusion] This study preliminarily indicates that Sophorae Tonkinensis is effective in playing the role of anti hepatoma through multiple components,multiple targets,and multiple pathways. This study provides a reference for elucidating the pharmacodynamic material basis and mechanism research of Sophorae Tonkinensis against liver cancer.