基于UPLC-Q-TOF-MS/MS和网络药理学探究六味安消颗粒治疗功能性便秘的药效物质基础及作用机制

To explore the pharmacodynamic material basis and mechanism of Liuwei Anxiao Keli(六味安消颗粒)in the treatment of functional constipation based on UPLC-Q-TOF-MS/MS and network pharmacology

目的探讨六味安消颗粒治疗功能性便秘(FC)的药效物质基础及作用机制。方法采用超高效液相色谱-四级杆-飞行时间串联质谱(UPLC-Q-TOF-MS/MS)技术分析六味安消颗粒的化学成分,并通过中药系统药理学数据库与分析平台(TCMSP)、DrugBank数据库筛选六味安消颗粒中各药物的活性成分;利用PubChem、Swiss Target Prediction数据库检索活性成分对应靶点,采用在线人类孟德尔遗传(OMIM)、Gene-Cards、DrugBank数据库检索FC疾病靶点,成分靶点与疾病靶点取交集后获得六味安消颗粒治疗FC的潜在靶点。利用Cytoscape 3.7.1软件构建“药物-成分-靶点”及蛋白质-蛋白质相互作用(PPI)网络并分析其核心成分及核心靶点。对潜在作用靶点进行基因本体(GO)富集分析及京都基因与基因组百科全书(KEGG)通路富集分析。利用Cytoscape 3.7.1软件构建“成分-靶点-通路”相互作用网络。利用AutodockVina.2.0对获得的核心成分及核心靶点进行分子对接验证。结果六味安消颗粒中共鉴定出化学成分61个;网络药理学分析结果显示,山柰酚、对甲氧基肉桂酸乙酯、大黄酸、异土木香内酯、鞣花酸、芦荟大黄素可能通过细胞肿瘤抗原(TP53)、原癌基因酪氨酸蛋白激酶(SRC)、丝氨酸/苏氨酸蛋白激酶1(AKT1)等核心靶点,调节磷脂酰肌醇-3-激酶-蛋白激酶B(PI3K-Akt)、丝裂原活化蛋白激酶(MAPK)等信号通路发挥治疗FC的作用;分子对接结果进一步证实上述核心成分与核心靶点间均具有良好的结合性能。结论该研究初步阐明了六味安消颗粒治疗FC的药效物质基础及作用机制,为后续的临床应用及药物开发提供了依据。

Objective To investigate the pharmacodynamic material basis and mechanism of Liuwei Anxiao Keli(六味安消颗粒)in the treatment of functional constipation(FC).Methods UPLC-Q-TOF-MS/MS technolo-gy was used to analyze the chemical components of Liuwei Anxiao Keli,and the active components and corresponding targets of each drug in Liuwei Anxiao Keli were screened by the Traditional Chinese Medicine System Pharmacology Database and analysis platform(TCMSP)and DrugBank database.PubChem and Swiss Target Prediction database were used to search the corresponding targets of active ingredients.Online human Mendelian Inheritance(OMIM),GeneCards and DrugBank databases were used to search FC disease targets,and the potential action targets of Liuwei Anxiao Keli in the treatment of FC were obtained after the intersection of component targets and disease targets.Cytoscape 3.7.1 was used to construct the“drug-component-target”and protein-protein interaction(PPI)network,and its core components and core targets were analyzed.Gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed for potential targets.Cytoscape 3.7.1 software was used to construct the“component-target-pathway”interaction network diagram.AutodockVina.2.0 was used to verify the molecular docking of the obtained core components and core targets.Results A total of 61 chemical components of Liuwei Anxiao Keli were identified.Network pharmacology analysis showed that kaempferol,ethyl p-methoxycinnamate,rhein,isoalantolactone,ellagaric acid and aloe emodin may act through core targets such as cellular tumor antigen(TP53),proto-oncogene tyrosine protein kinase(SRC)and serine/threonine protein kinase 1(AKT1)to regulate phosphatidylinositol-3-kinase-protein kinase B(PI3K-Akt),mitogen-activated protein kinase(MAPK)and other signaling pathways so as to play a therapeutic role in treatment of FC.The results of molecular docking further confirmed that the above core components have good binding properties with the core targets.Conclusion This study preliminarily illustrates the pharmacodynamic material basis and mechanism of Liuwei Anxiao Keli in the treatment of FC,and provides a basis for the subsequent clinical application and drug development.