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中国临床药理学与治疗学 ›› 2024, Vol. 29 ›› Issue (1): 11-25.doi: 10.12092/j.issn.1009-2501.2024.01.002

• 基础研究 • 上一篇    下一篇

基于UHPLC-Q-TOF/MS结合网络药理学与分子对接技术探究益心饮“异病同治”作用机制

王业健1,2,焦广洋3,庞  涛2,翁  楠4,高  洁2,李  娟1,陈万生2,3,陈卫东1,张  凤1,2   

  1. 1安徽中医药大学药学院,合肥  230012,安徽;
    2海军军医大学第二附属医院药剂科,上海  200003;
    3上海中医药大学中药研究所,上海  201203;
    4沈阳药科大学中药学院,辽宁  110000,沈阳

  • 收稿日期:2023-01-09 修回日期:2023-07-25 出版日期:2024-01-26 发布日期:2024-01-15
  • 通讯作者: 张凤,副教授,硕士生导师,研究方向:临床药学与中药学。 Tel: 021-51322403 E-mail: fengzhang@smmu.edu.cn 陈卫东,教授,博士生导师,研究方向:临床药学。 Tel: 0551-68129180 E-mail: wdchen@ahtcm.edu.cn
  • 作者简介:王业健,在读硕士,研究方向:中药药效物质基础研究。 E-mail:wyjjxhr@163.com
  • 基金资助:
    国家科技部重点研发项目(2022YFC3501700);海军军医大学附属长征医院人才建设三年行动计划“金字塔人才工程”(1016);上海市科学技术委员会项目(22S21901900)

Mechanism of Yi-xin-yin oral liquid according to homotherapy for heteropathy theory based on UHPLC-Q-TOF/MS combined with network pharmacology and molecular docking techniques

WANG Yejian1,2, JIAO Guangyang3, PANG Tao2, WENG Nan4, GAO Jie2, LI Juan1, CHEN Wansheng2,3, CHEN Weidong1, ZHANG Feng1,2   

  1. 1School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, Anhui, China; 2Department of Pharmacy, Changzheng Hospital, Naval Medical University (Second Military Medical University), Shanghai 200003, China; 3School of Pharmacy, Research and Development Center of Chinese Medicine Resources and Biotechnology, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 4School of Traditional Chinese Material, Shenyang Pharmaceutical University, Shenyang 110000, Liaoning, China
  • Received:2023-01-09 Revised:2023-07-25 Online:2024-01-26 Published:2024-01-15

摘要:

目的:根据中医“异病同治”理论,基于UHPLC-Q-TOF/MS技术、网络药理学、分子对接方法、细胞实验预测益心饮治疗心律失常、心力衰竭、心肌炎的核心靶点及相关信号通路。方法:采用UHPLC-Q-TOF/MS技术,对益心饮的化学成分和给药大鼠入血原形成分进行分析鉴定;并且通过TCMSP、SwissTargetPrediction等在线数据库获取入血原形成分潜在作用靶点,利用OMIM、Genecard等数据库获取疾病靶点,做Venn图获取交集靶点,并通过STRING11.5数据库构建蛋白质-蛋白质相互作用(PPI)网络,筛选核心靶点,并选用cytoscape3.9.0构建“疾病-成分-靶点”网络;利用DAVID数据库对核心靶点进行GO功能和KEGG富集分析,并运用Autodock vina软件进行分子对接验证,并以H9c2细胞对潜在活性成分和靶点进行验证。结果:从益心饮中鉴定得到157个化合物;从大鼠血清中鉴定得到34个化合物,主要包括姜辣素、异甘草素、甘草次酸等化合物,得到139个交集靶点,结合PPI网络筛选得到31个核心靶点,主要含有TNF、IL-6等靶点,KEGG通路富集分析主要涉及TNF信号通路、IL-17信号通路、MAPK信号通路、PI3K-Akt信号通路等。选取TNF、IL-6靶点与主要化合物进行分子对接,对接结果均小于-5 kcal/mol。体外细胞实验表明,益心饮能够通过调节TNF、IL-6发挥治疗作用。结论:益心饮主要潜在活性成分可能是异甘草素、甘草次酸、姜辣素、毛蕊异黄酮苷、丹酚酸B,主要通过作用于TNF、IL-6等靶点来调控特定的信号通路,发挥疗效。

关键词: 益心饮, 心力衰竭, 心律失常, 心肌炎, 异病同治, UHPLC-Q-TOF/MS, 网络药理学, 分子对接

Abstract:

AIM: To predict the core targets and related signaling pathways of Yi-xin-yin oral liquid for the treatment of arrhythmia, heart failure and myocarditis based on UHPLC-Q-TOF/MS, network pharmacology, molecular docking methods, cell experiments, according to the “homotherapy for heteropathy” theory in traditional Chinese medicine. METHODS: UHPLC-Q-TOF/MS was used to analyze and identify the chemical composition of Yi-xin-yin oral liquid Extract and the blood-absorbing components of rats oral administrated with Yi-xin-yin oral liquid extract, which compounds were applied in the databases searching for the potential targets (TCMSP, SwissTargetPrediction) and disease targets (OMIM, Genecard). Venn diagram was used for target intersection, and the subsequent protein-protein interaction network obtained core targets by STRING11.5 database, and then construct a "disease-component-target" network by cytoscape3.9.0. Finally, DAVID database was used to analysis GO function and KEGG enrichment analysis of core targets, and molecular docking validation was performed using Autodock vina software.And, validated with H9c2 cells for potential active ingredients and targets. RESULTS: A total of 156 compounds were identified from Yi-xin-yin Oral Liquid extract; 34 compounds were identified from rat serum, including 6-gingerol, isoliquiritigenin, glycyrrhizic acid and other compounds, and 139 intersecting targets were obtained. The KEGG pathway enrichment analysis mainly involved the TNF signaling pathway, IL-17 signaling pathway, MAPK signaling pathway, PI3K-Akt signaling pathway and so on. The TNF and IL-6 targets were selected for molecular docking with the main compounds, and the docking results were good (less than -5 kcal/mol). In vitro cellular experiments have shown that Yi-xin-yin oral liquid can exert therapeutic effects by regulating TNF and IL-6. CONCLUSION: The main potential active ingredients of Yi-xin-yin oral liquid may be isoliquiritigenin, glycyrrhetinic acid, calycosin-7-glucoside, salvianolic acid B, and 6-gingerol, which mainly act on TNF, IL-6 and other targets to regulate specific signaling pathways and exert therapeutic effects.

Key words: Yi-xin-yin oral liquid, heart failure, arrhythmia, myocarditis, homotherapy for heteropathy, UHPLC-Q-TOF/MS, network pharmacology, molecular docking

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