基于网络药理学及质谱技术的葶苈大枣泻肺汤抗心力衰竭的有效成分发现及作用机制分析

目的:基于网络药理学、高分辨质谱及分子对接技术探讨葶苈大枣泻肺汤治疗心力衰竭(简称心衰)的有效成分及作用机制。方法:通过文献查阅并结合BATMAN-TCM数据库对葶苈大枣泻肺汤中两味中药成分及药物靶标进行检索;以"heart failure"作为关键词,利用GeneCards数据库,对心衰疾病的靶标进行搜集;基于药物与疾病的共同靶标利用STRING数据平台进行蛋白质-蛋白质相互作用(PPI)分析,并通过Cytoscape软件进行网络拓补分析,最后得到核心靶标;基于核心靶标通过DAVID数据库进行京都基因与基因组百科全书(KEGG)通路富集分析,并建立"药物-成分-靶点-通路"分子网络。基于以上网络药理学研究结果,采用高分辨质谱技术对水煎汤剂进行分析,确证筛选出的活性成分;并采用AutoDock 4.2.6软件,对关键活性成分与相关靶标进行分子对接验证。结果:共获得葶苈子85种成分,大枣49种成分,药物靶标1 078个,疾病靶标1 549个,共同靶标262个。经蛋白质相互作用分析及网络拓补分析后,获得核心靶标23个,活性成分33个,涉及19条信号通路(P<0.05)。质谱分析结果表明异香草酸,南葶苈素A,山柰酚-7-O-β-D-吡喃葡萄糖苷等18个成分在水煎汤剂中得到确认;分子对接分析结果表示异香草酸,南葶苈素A,山柰酚-7-O-β-D-吡喃葡萄糖苷等6个核心成分(度值排名前6)与沉默信息调节因子1(SIRT1),白细胞介素(IL)1B,蛋白激酶Bα(AKT1),肿瘤坏死因子(TNF)有较好的结合活性;复方整体干预治疗心衰的过程主要涉及丝裂原活化蛋白激酶(MAPK),低氧诱导因子-1(HIF-1),雷帕霉素靶体蛋白(mTOR)等信号通路。结论:该研究初步揭示了葶苈大枣泻肺汤抗心衰的有效成分及其作用机制,对于指导临床精准用药及葶苈大枣泻肺汤质量控制标志物的筛选、抗心衰活性成分的发现提供参考。

Explore Effective Constituents and Mechanism of Tingli Dazao Xiefeitang in Treatment of Heart Failure Based on Network Pharmacology and Mass Spectrometry

Objective: To investigate the effective constituents and mechanism of Tingli Dazao Xiefeitang for the treatment of heart failure through network pharmacology and high resolution mass spectrometry technique as well as molecular docking technique. Method: Chemical components and potential targets in Descurainiae Semen Lepidii Semen and Jujubae Fructus were searched by referring to literature and BATMAN-TCM database. The disease targets were searched in Gene Cards database with ’’heart failure’’ as the key word. STRING platform was used to construct protein-protein interaction(PPI)network based on common targets of drugs and disease. The network topology was analyzed by using Cytoscape 3.7.2 software to obtain the core targets eventually. Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis of the core targets was conducted through DAVID database to draw a network of"herb-compound-target-pathway". Based on the results of the network pharmacology research above,high resolution mass spectrometry was used to analyze the decoction and confirm the selected active components. Auto Dock 4.2.6 software was used for molecular docking verification of key active components and related targets. Result: A total of 85 components were obtained in Descurainiae Semen Lepidii Semen,and 49 components were obtained in Fructus Ziziphi Jujubae. A total of 1 078 drug targets and 1 549 disease targets were identified. After PPI analysis and network topology analysis,23 core targets and 33 active components were obtained,involving 19 signaling pathways(P<0.05). Mass spectrometry analysis results indicated that 18 components such as isovanillic acid,descurainin A and kaempferol-7-O-β-D-glucopyranoside were confirmed in decoction. Molecular docking analysis results indicated that 6 core components(degree value top 6),such as isovanillic acid,descurainin A and kaempferol-7-O-β-Dglucopyranoside,had good binding activity with silent information regulatory factor 1(SIRT1),interleukin(IL)1 B,protein kinase Bα(AKT1)and tumor necrosis factor(TNF). The compound recipe for heart failure mainly involved mitogen-activated protein kinase(MAPK),hypoxia inducible factor-1(HIF-1),Mammalian target of rapamycin(m TOR) and other signaling pathways. Conclusion: This study preliminarily investigated the effective constituents and mechanism of Tingli Dazao Xiefeitang in the treatment of heart failure. It can provide references for the precise clinical medication and screening of quality control markers,as well as the discovery of active components in the treatment of heart failure.