基于UPLC-Q-Exactive Orbitrap-MS和网络药理学的芪参益气滴丸治疗心肌缺血的配伍机制研究

Compatibility mechanism of Qishen Yiqi Dripping Pills for treatment of myocardial ischemia based on UPLC-Q-Exactive Orbitrap-MS and network pharmacology

芪参益气滴丸(QSYQ)的临床疗效及作用机制已有较好研究基础,但发挥治疗作用的配伍机制还有待深入解析,该文旨在探究QSYQ治疗心肌缺血的配伍机制。采用UPLC-Q-Exactive Orbitrap-MS技术获得该方吸收入血成分;利用TCMSP、TCMIP和SwissTargetPrediction数据库收集并筛选入血成分的靶标蛋白;通过GeneCards、OMIM和DisGeNET数据库获得心肌缺血相关的疾病蛋白;利用在线绘图软件Venny 2.1.0、STRING和Cytoscape 3.9.1得到核心靶点与核心成分;通过DAVID数据库对核心靶点进行GO功能注释及KEGG通路富集,获得该方治疗心肌缺血的主要作用通路并绘制可视化网络图;基于“成分-靶点”“药材-通路”及“PI3K-AKT”特征通路分析配伍机制,并采用分子对接进行验证。该文获得QSYQ吸收入血成分42个,成分靶点556个,疾病靶点1980个,核心靶点69个,核心成分15个;该方可通过调控MAPK1、RELA、SRC、JUN、STAT3等蛋白,作用于HIF-1、PI3K-AKT、Toll-like、MAPK、VEGF等信号通路来发挥心肌缺血治疗功效;“成分-靶点”与“药材-通路”互作网络图在靶点与通路层面初步阐释了该方4味中药的协同情况;PI3K-AKT特征通路表明,君药黄芪与臣药丹参可调控该途径中的大部分靶点,佐药三七在IL6、AKT蛋白协同黄芪与丹参,使药降香在AKT、RXRA蛋白起调和作用,诸药共同作用于AKT、RXRA、NFKB等蛋白协同调控细胞的存活,进而起到促血管新生的作用。分子对接表明氢键和疏水作用可能是其主要作用形式,也验证了PI3K-AKT信号通路的结合能分布情况。该文从药材、成分、靶点、通路多维度解析该方的配伍内涵,为QSYQ作用机制及配伍规律的研究提供了参考依据。

Clinical efficacy and mechanism of Qishen Yiqi Dripping Pills(QSYQ) have been well researched, but the compatibility mechanism underlying its therapeutic effect still requires further analysis. This study aims to explore the compatibility mechanism of QSYQ in treating myocardial ischemia. UPLC-Q-Exactive Orbitrap-MS technique was used to obtain the absorbed blood components of QSYQ. Target proteins of the absorbed components were collected and screened using TCMSP, TCMIP, and SwissTargetPrediction databases. Disease proteins related to myocardial ischemia were obtained through GeneCards, OMIM, and DisGeNET databases. Core targets and core components were obtained using online plotting software Venny 2.1.0, STRING, and Cytoscape 3.9.1 software. David database was used for GO functional annotation and KEGG pathway enrichment of core targets, obtaining the main pathways of QSYQ in treating myocardial ischemia and drawing visualized network diagrams. The compatibility mechanism was analyzed based on "component-target", "drug-pathway", and "PI3K-AKT" characteristic pathways, and molecular docking was used for validation. This study obtained 42 absorbed blood components of QSYQ, 556 component targets, 1 980 disease targets, 69 core targets, and 15 core components. QSYQ can exert therapeutic effects on myocardial ischemia by regulating proteins such as MAPK1, RELA, SRC, JUN, and STAT3, acting on signaling pathways such as HIF-1, PI3K-AKT, Toll-like, MAPK, VEGF, etc. The interaction network diagrams of "component-target" and "drug-pathway" preliminarily elucidated the synergy among the four drugs in this prescription at the level of targets and pathways. The PI3K-AKT characteristic pathway indicated that the sovereign drug Huangqi(Astragali Radix) and minister drug Danshen(Salviae Miltiorrhizae Radix et Rhizoma) could regulate most targets in this pathway, while the assistant drug Sanqi(Notoginseng Radix et Rhizoma) cooperated with Huangqi and Danshen on IL6 and AKT proteins, and the envoy drug Jiangxiang(Dalbergiae Odoriferae Lignum) acted on AKT and RXRA proteins, with all drugs acting synergistically on proteins such as AKT, RXRA, NFKB to regulate cell survival and promote angiogenesis. Molecular docking indicated that hydrogen bonding and hydrophobic interactions might be the main forms of action, also validating the distribution of binding energy of the PI3K-AKT signaling pathway. This study analyzed the compatibility connotation of QSYQ from multiple dimensions including drugs, components, targets, and pathways, providing reference basis for the study of the mechanism of action and compatibility rules of QSYQ.