UPLC-Q-TOF-MS结合网络药理学探究参附汤治疗慢性心力衰竭的作用机制

Combination of UPLC-Q-TOF/MS and network pharmacology to explore the mechanism of Shenfu Decoction in the treatment of chronic heart failure

目的:通过UPLC-Q-TOF-MS技术检测在传统煎煮方法下参附汤的主要溶出成分,并结合网络药理学探究参附汤治疗慢性心力衰竭的作用靶点及机制。方法:采用UPLC-Q-TOF-MS技术对参附汤的主要溶出成分进行定性分析,并运用SwissTargetPrediction、PharmMapper数据库获得主要成分靶点;通过GeneCards、OMIM、DrugBank数据库收集慢性心力衰竭的相关治疗靶点;运用Venny 2.1构建韦恩图并得到交集靶点,结合STRING数据库构建靶蛋白相互作用网络,筛选核心靶点,并运用Cytoscape 3.9.1软件构建"化合物-靶点-疾病"网络;对药效疾病靶点进行GO功能和KEGG通路富集分析,并利用Atuodock软件对筛选核心靶点及活性成分进行分子对接技术验证关键成分与靶点蛋白的结合活性。结果:通过UPLC-Q-TOF-MS共鉴定参附汤主要成分39个,主要包含生物碱和皂苷类成分;疾病与活性成分交集靶点89个,其中包括血清白蛋白(ALB)、RAC-α丝氨酸/苏氨酸蛋白激酶(AKT1)、肿瘤坏死因子(TNF)、血管内皮生长因子A(VEGFA)、表皮生长因子受体(EGFR)等核心靶点;涉及的相关通路包括癌症、MAPK、cGMP-PKG、PI3K-Akt等信号通路;分子对接结果也进一步验证关键成分与核心靶点之间有较好结合能。结论:该研究通过UPLC-Q-TOF-MS结合网络药理学进一步阐明参附汤治疗慢性心衰主要靶点及通路,有助于了解其作用机制,为后续深入研究提供参考。

OBJECTIVE To detect the main soluble components of Shenfu Decoction by UPLC-Q-TOF-MS technology,and to explore the target and mechanism of Shenfu Decoction in the treatment of chronic heart failure combined with network pharmacology.METHODS The main dissolved components of Shenfu Decoction were qualitatively analyzed by UPLC-Q-TOF-MS technology,and the main component targets were obtained by SwissTargetPrediction and PharmMapper database.The related therapeutic targets of chronic heart failure were collected by GeneCards,OMIM and DrugBank databases,the Wayne diagram was constructed by Venny 2.1 and the intersection targets were obtained,the target protein interaction network was constructed by combining STRING database,the core targets were screened,and the"compound-target-disease"network was constructed by Cytoscape 3.9.1 software.The GO function and KEGG pathway enrichment analysis of drug-effective disease targets were carried out,and the molecular docking techniques were used to screen the core targets and active components to verify the binding activity of key components and target proteins.RESULTS A total of 39 main components of Shenfu Decoction were identified by UPLC-Q-TOF-MS,including alkaloids and saponins,and 89 intersection targets of disease and active components,including serum albumin(ALB),RAC-αserine/threonine protein kinase(Akt1),tumor necrosis factor(TNF),vascular endothelial growth factor A(VEGFA),epidermal growth factor receptor(EGFR)and so on.The related pathways involved include cancer,MAPK,cGMP-PKG,PI3K-Akt and other signal pathways;the results of molecular docking also further verify that there is a good binding energy between the key components and the core target.CONCLUSION This study further elucidates the main targets and pathways of Shenfu Decoction in the treatment of chronic heart failure through UPLC-Q-TOF-MS combined with network pharmacology,which is helpful to understand its mechanism and provide reference for further research.