GC-MS、UPLC-Q-TOF-MS/MS结合网络药理学探讨地锦草药效物质基础及其抗炎作用机制

The therapeutic material basis of Dijincao(Euphorbiae Humifusae Herba)and its anti-inflammatory mechanism based on GC-MS,UPLC-Q-TOF-MS/MS,and network pharmacology

目的采用色谱联用技术分析地锦草主要成分,并联合网络药理学预测其抗炎作用的潜在靶点和信号通路,阐明地锦草治疗炎症的作用机制。方法通过GC-MS、UPLC-Q-TOF-MS/MS技术分析地锦草的化学成分,借助中药系统药理数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology,TCMSP)、地锦草相关文献筛选地锦草活性成分;采用SwissTargetPrediction数据库预测靶点,采用GeneCards、DrugBank、DisGeNET数据库获取地锦草相关治疗靶点,将成分靶点和疾病靶点取交集,获得交集靶点;凭借STRING 11.5数据库和Cytoscape 3.9.0软件构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并筛选核心靶点。采用DAVID 6.8进行GO注释和KEGG通路富集分析,构建“化合物-靶点-通路”网络。结果GC-MS、UPLC-Q-TOF-MS/MS分别鉴定出50、76种成分,筛选出地锦草抗炎的12个活性成分及214个成分靶点;根据数据库筛选得到10688个抗炎相关靶点、活性成分与疾病交集靶点198个;借助CytoNCA插件筛选出核心靶点37个;DAVID数据库富集分析显示,地锦草主要通过磷脂酰肌-3-激酶-蛋白激酶B(phosphatidylinositol-3-kinase-protein kinase B,PI3K/PKB)、酪氨酸激酶受体、血管内皮生长因子(vascular endothelial growth factor,VEGF)、甲状腺激素、雌激素等信号通路发挥治疗炎症的作用。结论地锦草通过多成分、多靶点、多途径发挥治疗炎症的作用,体现了中药复杂系统的作用特点,为全面阐释地锦草药效物质基础提供一定参考。

Objective To elucidate the mechanism of action of Dijincao(Euphorbiae Humifusae Herba)in treating inflammation through analyzing the main components of Dijincao(Euphorbiae Humifusae Herba)by chromatographic coupling techniques combined with predicting the potential targets of action and signaling pathways for anti-inflammation by network pharmacology.Methods The chemical composition of Dijincao(Euphorbiae Humifusae Herba)were analyzed by GC-MS and UPLC-Q-TOF-MS/MS techniques and the active ingredients were screened by Traditional Chinese Medicine Systems Pharmacology(TCMSP)and related literature.The targets were predicted by Swiss Target Prediction and the Dijincao(Euphorbiae Humifusae Herba)-related therapeutic targets were obtained by the GeneCards,DrugBank,and DisGeNET.Then the component targets and disease targets were intersected to obtain intersecting targets.The STRING 11.5 database and Cytoscape 3.9.0 software were used to construct the protein-protein interaction(PPI)networks and screen the core targets.GO annotation and KEGG pathway enrichment analysis were performed by DAVID 6.8 to construct a"compound-target-pathway"network.Results A total of 50 and 76 components were identified by GC-MS and UPLC-Q-TOF-MS/MS respectively,and 12 active components and 214 component targets were screened.The total of 10688 anti-inflammatory-related targets and 198 active component-disease intersection targets were obtained based on database screening and 37 core targets were screened by CytoNCA plug-in.The enrichment analysis of DAVID database showed that Dijincao(Euphorbiae Humifusae Herba)mainly exerted its therapeutic effects on inflammation through signaling pathways of phosphatidylinositide 3-kinases-protein kinase B(PI3K-Akt),ErbB,vascular endothelial growth factor(VEGF),thyroid hormone,estrogen,and so on.Conclusion Dijincao(Euphorbiae Humifusae Herba)exerts its therapeutic effects on inflammation through multiple components,multiple targets,and multiple pathways,which is the embodiment of the action characteristics of the complex system of Chinese medicines.It also provides a reference for the comprehensive elucidation of therapeutic material basis of Dijincao(Euphorbiae Humifusae Herba).