摘要
目的基于网络药理学分析活络效灵丹治疗冠心病的分子机制,并通过分子对接及动物实验对其进行验证。方法使用TCMSP2.3收集活络效灵丹中四味中药的化学成分和相关靶点,使用DisGeNET、DrugBank、GeneCards和OMIM数据库搜索冠心病的疾病靶点,将药物靶点与疾病靶点取交集作为共有靶点,在STRING数据平台中导入共有靶点获得蛋白-蛋白相互作用的PPI网络关系。将共有靶点上传至DAVID6.8数据库进行生物富集分析,使用Cytoscape3.7.1分析软件构建PPI网络图与“成分-靶点-通路”网络图,并利用分子对接技术验证关键靶点与相关化合物的结合程度。通过建立大鼠心肌缺血模型,采用HE染色与Masson染色检测心肌组织病理结果;采用免疫荧光与Western blot法验证活络效灵丹对PTGS2表达的影响。结果通过筛选获得活络效灵丹有效成分83个,相关靶点216个,疾病相关靶点695个,共有靶点50个。GO生物富集分析结果显示共有靶点主要涉及调控凋亡过程、正向调节细胞增殖、炎症反应、应对缺氧等。KEGG通路共富集到76条通路,主要与TNF、PI3K-Akt、NF-κB、HIF-1和MAPK信号通路等有关。分子对接验证显示核心靶点与核心化合物具有较好的结合活性。动物实验证明活络效灵丹能够明显的改善心肌损伤,免疫荧光与Western blot证明其能够抑制心肌组织中PTGS2的表达。结论本次研究将网络药理学与分子对接及实验验证相结合,明确了活络效灵丹治疗冠心病的有效成分、潜在靶点和作用机制,为其临床应用提供了依据。
Objective To determine the molecular mechanism of Huoluo Xiaoling pill(HLXLP)for coronary heart disease(CHD)based on network pharmacology,and to verify the molecular mechanism of HLXLP in the treatment of CHD by experiments and molecular docking.Methods Firstly,constituents and targets of HLXLP were collected with TCMSP2.3,and the disease targets of CHD were obtained from DisGeNET,DrugBank,GeneCards and OMIM databases.Then the common targets were obtained by integrating the drug targets and disease targets,which imported into the online platform of STRING to obtain the protein-protein interaction(PPI).GO and KEGG enrichment analysis of the common targets were conducted through DAVID6.8.The“component-target-pathway”and PPI networks were constructed with Cytoscape3.7.1.Finally,the main active components and key targets of HLXLP were verified by molecular docking with Autodock vina.The myocardial ischemia rat models were established,and the expression of PTGS2,and histopathological changes in the myocardium were investigated.Results Totally 83 active components,216 targets for HLXLP,695 targets for CHD,and 50 common targets were obtained.GO analysis showed that the target genes were mainly related to biological processes such as negative regulation of apoptotic process,positive regulation of cell proliferation,inflammatory response and response to hypoxia.A total of 76 signal pathways were obtained by KEGG pathway enrichment analysis,which mainly included TNF,PI3K-Akt,NF-κB,HIF-1 and MAPK related signaling pathways.The molecular docking revealed that the active components and targets showed good binding interactions.The HE staining and Masson staining showed that HLXLP significantly improved the myocardial injury,and Western blot and Immunofluorescence showed that HLXLP inhibited the expression of PTGS2.Conclusion This study combines network pharmacology with molecular docking and experimental verification to clarify the active ingredients,potential targets and mechanism of HLXLP for CHD,providing a basis for its clinical application.
作者
赵佳鑫
张娟利
马阳
黄少杰
陈海霞
牟菲
文爱东
丁一
ZHAO Jia-xin;ZHANG Juan-li;MA Yang;HUANG Shao-jie;CHEN Hai-xia;MU Fei;WEN Ai-dong;DING Yi(College of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046;Department of Pharmacy,The First Affiliated Hospital of Air Force Military Medical University,Xi’an 710032)
出处
《中南药学》
CAS
2022年第3期565-573,共9页
Central South Pharmacy
基金
国家自然科学基金项目(No.82074321
No.81903837)
陕西省重点研发计划(No.2021KW-56)。
关键词
网络药理学
活络效灵丹
分子对接
冠心病
network pharmacology
Huoluo Xiaoling pill
molecular docking
coronary heart disease
