基于UPLC-Q-TOF-MS/MS和网络药理学结合分子对接的凹叶景天抗肝癌质量标志物(Q-Marker)研究

Study on anti-liver cancer quality markers(Q-Marker)of Sedum emarginatum based on UPLC-Q-TOF-MS/MS and network pharmacology combined with molecular docking

目的 利用高效液相色谱-四极杆飞行时间串联质谱法(UPLC-Q-TOF-MS/MS)分析凹叶景天醋酸乙酯部位的化学成分,并结合网络药理学进行凹叶景天抗肝癌质量标志物(Q-Marker)的预测分析。方法 利用UPLC-Q-TOF-MS/MS快速鉴定凹叶景天醋酸乙酯部位的化学成分;利用Swiss ADME平台,根据筛选原则确定活性成分;运用Swiss Target Prediction平台预测凹叶景天的成分靶点,GeneCards平台获得相关疾病靶点,Venny平台获得成分和疾病的交集靶点,并构建“活性成分-靶点”网络;通过String数据库和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)网络,并筛选核心靶点;利用David数据库对潜在的核心靶点进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析;采用分子对接技术进行验证。结果 从凹叶景天醋酸乙酯部位中共鉴定58个成分,黄酮类23个、有机酸类16个、酚类4个、香豆素类4个、萜类2个、苯酞类2个等。网络药理学研究筛选出39种活性成分,药物-疾病靶点140个;PPI网络筛选出AKT1、EGFR、STAT3、CASP3、ESR1等18个核心靶点;富集分析显示,凹叶景天主要以癌症通路、PI3K-Akt信号通路、癌症中的微小RNA信号通路、MAPK信号通路等为主要信号通路,从而发挥抗肝癌的作用。分子对接结果显示预测的QMarker及核心靶点之间具有良好的亲和力。结论 凹叶景天是通过多成分、多靶点、多途径发挥抗肝癌的作用,预测出山柰素、异鼠李素、山柰酚、槲皮素、木犀草素、芹菜素和苜蓿素7个成分可作为凹叶景天抗肝癌的Q-Marker。

Objective UPLC-Q-TOF-MS/MS was used to analyze the chemical composition of ethyl acetate in Sedum emarginatum,and the predictive analysis of quality markers(Q-Marker)of anti-liver cancer in S.emarginatum was carried out combined with network pharmacology.Methods The chemical constituents of ethyl acetate fraction of S.emarginatum were rapidly identified by UPLC-Q-TOF-MS/MS.According to the screening principle,the active components were determined by Swiss ADME platform.The Swiss Target Prediction platform was used to predict the component targets of S.emarginatum.The GeneCards platform was used to obtain the related disease targets.The Venny platform was used to obtain the intersection targets of components and diseases,and the'active component-target'network was constructed.The protein-protein interaction(PPI)network was constructed by String database and Cytoscape software,and the core targets were screened.Gene Ontology(GO)function enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)signal pathway enrichment analysis of potential core targets were performed using the David database.Molecular docking technology was used for verification.Results A total of 58 components were identified from the ethyl acetate extract,including 23 flavonoids,16 organic acids,four phenols,four coumarins,two terpenoids,and two phthalides etc.A total of 39 active components and 140 drug-disease targets were screened out by network pharmacology.Eighteen core targets such as AKT1,EGFR,STAT3,CASP3 and ESR1 were screened from PPI network.Enrichment analysis showed that S.emarginatumm played an anti-liver cancer role mainly through cancer pathway,PI3K-Akt signaling pathway,microRNA signaling pathway in cancer and MAPK signaling pathway,etc.Molecular docking results showed that there was a good affinity between the predicted Q-Marker and the core targets.Conclusion This study preliminarily clarified that S.emarginatum played an anti-liver cancer role through multi-component,multi-target and multi-pathway.It was predicted that kaempferide,isorhamnetin,kaempferol,quercetin,luteolin,apigenin and tricin could be used as Q-Marker for anti−liver cancer of S.emarginatum.