基于UHPLC/Q-Orbitrap-MS的石决明散化学成分鉴定及降压作用网络药理学研究

Identification of chemical compounds of Shi-Jue-Ming-San based on ultra-high performance liquid chromatography-quadrupole Orbitrap mass spectrometry and its network pharmacology in treatment of hypertension

关键词:  石决明散  化学成分  高血压  网络药理学  作用机制

Key words:  Shi-Jue-Ming-San  chemical composition  hypertension  network pharmacology  mechanism of action

摘要:

[目的] 基于入血成分分析和网络药理学探讨石决明散（SJMS）抗高血压（HT）的潜在作用机制。[方法] 利用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱联用（UHPLC/Q-Orbitrap-MS）技术对SJMS中的主要化学成分和入血成分进行定性分析，利用SEA和SwissTargetPrediction数据库预测入血成分的作用靶点；借助GeneCards和DrugBank数据库获得HT的相关靶点；通过String平台和Cytoscape软件对SJMS抗HT的潜在靶点进行PPI网络构建；使用DAVID数据库对交集靶点进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）代谢通路富集分析；最后采用Cytoscape软件构建“成分-疾病-通路-靶点”网络图分析。[结果] 方剂共鉴定出129个主要化学成分和23个原型入血成分；获得SJMS抗HT的交集靶点共296个，其中SRC、TP53、STAT3、EGFR、AKT1、ESR1等是SJMS抗HT的关键作用靶点；橙黄决明素、红链霉素-6-O-β龙胆二糖苷、决明柯酮、甘草次酸、新绿原酸、甘草酸、芹菜素等为SJMS抗HT的主要活性成分；GO富集到基因功能30个条目，差异具有统计学意义（P<0.01），SJMS抗HT与蛋白质磷酸化、凋亡过程的负调控、细胞增殖的正向调节等有关；KEGG通路共20条，差异具有统计学意义（P<0.01），结果显示：SJMS主要通过缺氧诱导因子-1（HIF-1）信号通路、高级糖基化终末产物-受体（AGE-RAGE）信号通路等来发挥治疗HT的作用。[结论] 研究初步阐明了SJMS的潜在药效物质基础，探讨了SJMS抗HT的潜在作用机制，为其临床应用提供了一定的理论参考。

Abstract:

[Objective] Based on the analysis of absorbed components in blood and network pharmacology,the potential mechanism of Shi-Jue-Ming-San(SJMS) in treating hypertension(HT) was explored. [Methods] Qualitative analysis was conducted on the main chemical compounds in vivo and in vitro of SJMS using the ultra-high performance liquid chromatography-quadrupole Orbitrap mass spectrometry(UHPLC/Q-Orbitrap-MS),and the targeted protein of active compounds were predicted using SEA and SwissTargetPrediction databases; Using GeneCards and DrugBank databases to obtain relevant targets for hypertension; Construct a PPI network for potential targets of SJMS on HT using the String platform and Cytoscape software; Perform GO functional enrichment analysis and KEGG metabolic pathway enrichment analysis on intersection targets using the DAVID database; Finally, a "component-disease-pathway-target" network diagram was constructed using Cytoscape software for analysis. [Results] 129 main chemical compounds and 23 prototype components in blood were identified in SJMS; A total of 296 intersection targets were obtained,among which SRC,TP53,STAT3,EGFR,AKT1,ESR1 are key targets for SJMS anti-HT. The main active ingredients of SJMS for anti-HT include aurantio-obtusin,rubrofusarin-6-O-β-gentiobioside,torachrysone,glycyrrhetinic acid,neochlorogenic acid,glycyrrhizictinic acid,apigenin; GO enriched 30 gene function entries(P<0.01),and the activity of SJMS was negatively related to protein phosphorylation,and apoptosis process,while positively related to cell proliferation. There were 20 of KEGG pathways(P<0.01). The results showed that SJMS played a role in treating HT mainly through HIF-1 signaling pathway and AGE-RAGE signaling pathway in diabetes complications. [Conclusion] This study preliminarily elucidates the potential pharmacological substance basis of SJMS and explores its potential mechanism of action against HT,providing a certain theoretical reference for its clinical application.