血清药物化学结合网络药理学探讨养心氏片治疗冠心病的物质基础与作用机制

Combining Serum Pharmacochemistry and Network Pharmacology to Explore the Pharmacodynamic Material Basis and Mechanisms of Yangxinshi Tablet in Treating Coronary Heart Disease

关键词:  养心氏片  冠心病  入血成分  网络分析  炎症反应  氧化应激

Key words:  Yangxinshi tablet  coronary heart disease  prototype compound  network analysis  inflammatory response  oxidative stress

摘要:

目的 探讨养心氏片治疗冠心病的药效物质基础与潜在的作用机制。方法 采用HPLC-ESI-Q-TOF-MS/MS技术对养心氏片（Yangxinshi tablet，YXST）的入血成分进行分析；利用STITCH、TCMSP、Binding DB数据库预测YXST入血成分的靶点；采用TTD、DrugBank、PharmGKB、MalaCards、CTD数据库收集冠心病相关靶点；通过STRING数据库对得到的成分靶点和疾病靶点构建蛋白相互作用关系（protein-protein interaction，PPI）网络从而筛选YXST治疗冠心病的潜在靶点；利用R软件的clusterProfiler包对潜在靶点进行基因本体论（gene ontology，GO）功能注释和京都基因和基因组百科全书（kyoto encyclopedia of genes and genomes，KEGG）通路分析；采用Cytoscape软件构建"入血成分-潜在靶点"网络并分析关键化合物和关键靶点。结果 通过对大鼠口服灌胃给予YXST提取液后的血浆进行分析，共检测到26个原型成分。在数据库中收集得到入血成分对应的287个靶点和冠心病相关的199个靶点。通过交集分析和PPI网络的构建共获得70个YXST治疗冠心病的潜在靶点。对潜在靶点进行GO功能注释和KEGG通路分析发现，YXST治疗冠心病主要涉及了炎症反应、氧化应激以及晚期糖基化终末产物-晚期糖基化终末产物受体（advanced glycation end products-receptor for advanced glycation end products，AGE-RAGE）、动脉粥样硬化、肿瘤坏死因子（tumour necrosis factor，TNF）、白细胞介素-17（interleukin-17，IL-17）、C型凝集素受体、Th17细胞分化、Toll样受体等信号通路。通过构建"入血成分-潜在靶点"网络分析得到3个关键化合物和16个关键靶点。结论 YXST可通过调节TNF、PTGS2、NOS2、NOS3等关键靶点干预炎症反应、氧化应激、动脉粥样硬化等过程发挥治疗冠心病的作用，其潜在的有效成分为大豆苷、小檗碱和葛根素。本研究充分体现了YXST通过多成分-多靶点-多途径治疗冠心病的作用特点，为深入研究YXST的药理作用机制提供重要参考。

Abstract:

OBJECTIVE To explore the pharmacodynamic material basis and mechanisms of Yangxinshi tablet(YXST) in treating coronary heart disease. METHODS HPLC-ESI-Q-TOF-MS/MS method was performed to analyze the blood-entry components of YXST. The targets of blood-entry compounds were derived from STITCH, TCMSP and Binding DB database. The targets related to coronary heart disease were collected from TTD, DrugBank, PharmGKB, MalaCards and CTD database. Protein-protein interaction(PPI) network was constructed for the obtained compound targets and disease targets using STRING database to screen potential anti-coronary heart disease targets of YXST. Gene ontology(GO) function annotation and kyoto encyclopedia of genes and genomes(KEGG) pathway analysis of potential targets were performed using the clusterProfiler package in R. The blood-entry components-potential target network was further constructed by Cytoscape software to obtain key compounds and key targets. RESULTS Rat plasma after oral administration of YXST was analyzed and 26 prototype compounds could be detected. A total of 287 compound targets corresponding to blood entry components and 199 disease targets associated with coronary heart disease were collected in the database. By intersection analysis and PPI network construction, 70 potential anti-coronary heart disease targets of YXST were obtained. According to the results of GO function annotation and KEGG pathway analysis, YXST treating coronary heart disease mainly involved in the inflammatory response, oxidative stress and advanced glycationend products-receptor for advanced glycationend products(AGE-RAGE), atherosclerosis, tumour necrosis factor(TNF), interleukin-17(IL-17), C-type lectin receptor, Th17 cell differentiation and Toll-like receptor signaling pathways. By constructing and analyzing the blood-entry components-potential target network, 3 key compounds and 16 key targets were obtained. CONCLUSION YXST can interfere with the inflammatory response, oxidative stress and atherosclerosis process by regulating TNF, PTGS2, NOS2 and NOS3. The potential effective compounds are daidzein, berberine and puerarin. This study fully reflect the characteristics of YXST in treating coronary heart disease through multi-component, multi-target and multi-pathway, and provide an important reference for further research on the pharmacological mechanisms of YXST.