摘要
目的基于网络药理学探讨银丹心脑通软胶囊(YD)治疗心肌缺血再灌注损伤(MIRI)的物质基础、作用机制和实验验证,为该药治疗MIRI提供理论参考。方法利用TCMSP、Swiss Target Prediction、中国知网等数据库检索YD的主要活性成分和作用靶点,GeneCards数据库筛选MIRI疾病靶点,Cytoscape软件构建“药物-成分-疾病-靶点”网络关系,富集分析预测作用机制,并将排名前三的主要活性成分与PPI网络中排名前六的靶点进行进行分子对接验证。人心肌AC-16细胞进行低氧/复氧(H/R)模型构建,初步验证核心靶点。通过CCK-8比色法检测细胞活力,探索最适药物浓度。光镜下观察银丹心脑通对心肌细胞形态学变化影响。LDH渗漏检测细胞膜完整性;Western blot检测STAT3、p-STAT3、PI3K、AKT1和p-AKT1的表达。结果共得到YD活性成分105个,药物靶点382个,MIRI疾病靶点1223个,药物-疾病共同靶点160个,关键靶点涉及AKT1、STAT3、VEGFA、TNF、MAPK8、PIK3CA等,GO析主要涉及细胞凋亡、脂溶反应、肌肉细胞增殖、细胞因子介导的炎症反应、氧化应激等。分子对接结果显示,VEGFA、APP、PIK3CA与槲皮素、木犀草素、山柰酚都能较好的结合。细胞实验显示,200 mg/L的YD能显著地促进AC-16细胞增殖,减少LDH渗漏。Western blot结果显示,银丹心脑通能激活STAT3和PI3K-AKT1信号通路,保护心肌。结论YD能通过多成分、多靶点、多通路保护MIRI。
Objective To explore the material basis and mechanism of Yindan Xinnaotong soft capsule(YD)in treating myocardial ischemia-reperfusion injury(MIRI)based on network pharmacology,so as to provide theoretical reference for the treatment of MIRI.Methods The main active components and action targets of YD were searched by TCMSP,SwissTargetPrediction and China knowledge Network.The disease targets of MIRI were screened by GeneCards database.The“drug-component-disease-target”network relationship was constructed by Cytoscape software,enrichment analysis and prediction mechanism were analyzed,and the molecular docking verification was carried out between the top three main active components and the top six targets in PPI network.Human myocardial AC-16 cells were constructed hypoxia/reoxygenation(H/R)model to initially verify the core target.Using CCK-8 assay was usedto detect cell viability and explore the optimal drugconcentration.The influence of YD on cell morphology was observed by using optical microscope.LDH content was detected to assess the integrity of cell membrane;Western blot was used to detect the expression of STAT3,p-STAT3,PI3K,AKT1 and p-AKT1.Results A total of 105 active components,382 drug targets,1223 MIRI disease targets and 160 drug-disease common targets of YD were obtained.The key targets involved AKT1,STAT3,VEGFA,TNF,MAPK8,PIK3CA and so on.GO analysis mainly involved apoptosis,lipolysis,muscle cell proliferation,cytokine-mediated inflammation,oxidative stress and so on.The results of molecular docking showed that VEGFA,APP and PIK3CA could bind to quercetin,luteolin and kaempferol.Our results showed that 200 mg/L YD could significantly promote the proliferation of AC-16 cells and reduce LDH leakage.Western blot results showed that YD could activate STAT3 and PI3K-AKT1 signaling pathways and protect myocardium.Conclusion YD can protect MIRI through multi-components,multi-targets and multi-pathways.
作者
徐启丽
邹常超
莫丽莉
周海燕
刘兴德
Xu Qili;Zou Changchao;Mo Lili;Zhou Haiyan;Liu Xingde(Dept of Cardiology, Affiliated Hospital of Guizhou Medical University,Guiyang 550000;Dept of Cardiology, Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550000)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第1期131-138,共8页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:82060855、31760294)
贵州省科技计划项目(编号:黔科合平台人才[2018]5608)
贵阳市科技计划项目:(编号:筑科合同[2017]30-10)
贵州省教育厅青年科技人才成长项目:(编号:黔教合KY字[2018]182)。
