摘要
目的对冠心Ⅱ号方的活性成分进行鉴定,探讨其在治疗急性心肌梗死中作用机制。方法采用高效液相色谱-质谱联用技术(HPLC-MS/MS)对冠心Ⅱ号方的成分进行鉴定;利用中药系统药理数据库筛选出已检测出成分的作用靶点;利用GendCards、OMIM数据库筛选疾病相关靶点;使用STRING平台对靶点进行PPI网络构建;利用Bioconductor数据库对靶点进行GO和KEGG富集分析;利用Cytoscape 3.7.2软件构建中药-成分-靶点-通路-疾病网络图,然后通过拓扑学参数分析获得冠心Ⅱ号方治疗急性心肌梗死的核心靶点,最后将鉴定出的质控成分与核心靶点进行分子对接验证。结果整合HPLC-MS/MS和网络药理学结果,检测出冠心Ⅱ号方5种化学成分,分别为阿魏酸、芍药苷、羟基红花黄色素A、丹参素、原儿茶醛;筛选出CHRM1、CHRM2、CHRM3、PTGS1、ADRA1A、PLAU、GABRA1、AR、IL6共9个关键靶点,涉及神经活性配体受体相互作用、钙信号通路、胆碱能突触、肌动蛋白细胞骨架调节4条信号通路。分子对接显示,5种质控成分与核心靶点CHRM1具有较好的结合活性,分子构象结合稳定。结论HPLC-MS/MS可快速、准确检测出冠心Ⅱ号方5种质控成分。联合网络药理学研究对鉴定出的5种成分进行靶点与通路分析,冠心Ⅱ号方5种质控成分可通过与CHRM1靶蛋白结合,调节神经活性配体受体相互作用、钙信号通路、胆碱能突触、肌动蛋白细胞骨架调节信号通路来抑制心梗后的氧化应激、炎症损伤和细胞凋亡,发挥保护心肌细胞的作用。本研究为进一步阐述冠心Ⅱ号方在急性心肌梗死中的作用机制提供了新的思路和方向。但是,冠心Ⅱ号方是否通过上述4条通路发挥作用及通路间的主要作用靶点和通路间相互情况仍需要进一步的实验证明。
Objective To identify the chemical components of GuanxinⅡformula,and explore its mechanism in the treatment of acute myocardial infarction.Methods In this study,the components of GuanxinⅡformula were identified with high performance liquid chromatography-mass spectrometry-tandem mass spectrometry(HPLC-MS/MS),the targets of the detected components were screened by searching the TCMSP database,and the disease targets were screened by searching the GeneCards and OMIM databases.Then STRING was applied to build the network of PPI,and Bioconductor database was used to perform GO and KEGG enrichment analysis,and Cytoscape software was used to build the drug-component-target-pathway-disease network diagram.Core targets were then obtained by calculating topological parameters.Finally,detected components and targets were identified by molecular docking.Results Five chemical components of GuanxinⅡformula were detected,i.e.ferulic acid,tanshinol,paeoniflorin,protocatechualdehyde,and hydroxysafflor yellow A(HSYA).Nine key targets were screened,including CHRM1,CHRM2,CHRM3,PTGS1,ADRA1 A,PLAU,GABRA1,AR and IL6,with the neuroactive ligand-receptor interaction pathway,calcium signaling pathway,cholinergic synapse pathway,and actin cytoskeleton signaling pathway involved.Molecular docking showed good binding activity between the detected components and CHRM1,and the molecular conformation was stable.Conclusion Five quality control components of GuanxinⅡformula could be detected relatively quickly and accurately by HPLC-MS/MS.By binding with CHRM1,the components of GuanxinⅡseem to protect cardiac myocytes through regulating the neuroactive ligand-receptor interaction pathway,calcium signaling pathway,cholinergic synapse pathway,and actin cytoskeleton signaling pathway,thus inhibiting oxidative stress,inflammatory damage and apoptosis after myocardial infarction.This study potentially provides a new idea and direction for further exploring the mechanism of GuanxinⅡformula in the treatment of acute myocardial infarction.However,further experimental proof is still needed to determine whether GuanxinⅡformula plays a role through the four pathways above,and to clarify the main targets and interaction between pathways.
作者
冯娟
闫奎坡
朱翠玲
朱明军
孙彦琴
徐亚洲
郭雨晴
刘晓萌
Feng Juan;Yan Kuipo;Zhu Cuiling;Zhu Mingjun;Sun Yanqin;Xu Yazhou;Guo Yuqing;Liu Xiaomeng(Henan University of Chinese Medicine,Henan 450000,China;The First AHilialed Hospital of Henan University of Chinese Medicine,Henan 450000,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2021年第8期704-713,共10页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家重点研发计划项目(No.2019YFC1710000,No.2019YFC1710001,No.2019YFC1710003)
国家自然科学基金青年基金项目(No.81303073)
河南省首批青苗人才培养项目[豫中医科教(2018)No.16]
河南省科技攻关项目(No.192102310161,No.182102310291)
河南省中医药科学研究专项(No.2016ZY3003,No.2021ZY1015,No.2021ZY2004)。
