基于网络药理学和斑马鱼模型的心可舒片抗心肌缺血活性成分及作用机制研究  

Study on anti-myocardial ischemia active components and mechanism of Xinkeshu tablets based on network pharmacology and zebrafish model

作　　者：侯林华 张华铮 高硕 张云 何秋霞 刘可春 孙晨 李建恒[1] 夏青 HOU Lin-hua;ZHANG Hua-zheng;GAO Shuo;ZHANG Yun;HE Qiu-xia;LIU Ke-chun;SUN Chen;LI Jian-heng;XIA Qing(College of Pharmaceutical Sciences,Hebei University,Baoding,Hebei 071002,China;Qilu University of Technology(Shandong Academy of Sciences),Biology Institute of Shandong Academy of Sciences,Jinan 250103,China;Shandong Academy of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]河北大学药学院,河北保定071002 [2]齐鲁工业大学(山东省科学院),山东省科学院生物研究所,山东济南250103 [3]山东省中医药研究院,山东济南250014

出　　处：《中国药理学通报》2024年第5期964-974,共11页Chinese Pharmacological Bulletin

基　　金：国家重点研发计划(No 2022YFC2804600);山东省自然科学基金项目(No ZR2021QH155);山东省中医药科技项目(No 2021Q124);山东省中央引导地方科技发展资金项目(No YDZX2022164);山东省科技型中小企业创新能力提升工程项目(No 2022TSGC2003);“药物制剂技术研究与评价”国家药品监督管理局重点实验室开放课题(No 2021TREDP01)。

摘　　要：目的基于网络药理学和斑马鱼模型研究心可舒片抗心肌缺血活性成分及作用机制。方法利用盐酸异丙肾上腺素(isoprenaline,ISO)诱导的斑马鱼心肌缺血模型和过氧化氢(H_(2)O_(2))诱导的大鼠心肌细胞(H9c2)损伤模型,评价心可舒片抗心肌缺血活性。利用TCMSP等数据库检索心可舒片的活性成分;利用PharmMapper数据库预测潜在作用靶点;利用OMIM数据库检索心肌缺血疾病靶点;分析心可舒片抗心肌缺血的潜在治疗靶点;对核心靶点进行GO和KEGG富集分析。利用斑马鱼和细胞模型对活性成分进行验证;利用qRT-PCR检测心可舒片对关键靶点表达的影响。结果心可舒片可以明显缓解ISO诱导的斑马鱼心包水肿、心动过缓、静脉窦-动脉球(SV-BA)距离增大,提高细胞存活率。心可舒片的30个潜在活性成分主要作用于ALB、AKT1、MAPK1等30个核心靶点;通过调节蛋白质磷酸化、负调控凋亡、正调控PI3K信号转导等627个GO条目,调控PI3K/Akt、FOXO、Ras等117条信号通路发挥抗心肌缺血作用。丹酚酸A、紫草酸、迷迭香酸、丹酚酸D、丹酚酸B、人参皂苷Rg2、金丝桃苷、3′-甲氧基葛根素、3′-羟基葛根素和人参皂苷Rg1能够缓解ISO引起的斑马鱼心肌缺血、提高缺氧细胞存活率。心可舒片能够上调ras、akt1等基因的表达,下调mapk1、mapk8等基因的表达。结论心可舒片中的丹酚酸A、紫草酸等活性成分通过靶向AKT1、MAPK1等靶点,调控PI3K/Akt、MAPK、Ras等信号通路发挥抗心肌缺血作用。Aim To study the active ingredients and mechanism of action of Xinkeshu tablets against myocardial ischemia by network pharmacology and zebrafish model.Methods The anti-myocardial ischemia activity of Xinkeshu tablets was evaluated by isoprenaline hydrochloride(ISO)-induced zebrafish myocardial ischemia model and H_(2)O_(2)-induced H9c2 damage model.The active ingredients of Xinkeshu tablets were retrieved using databases such as TCMSP.The potential targets were predicted by PharmaMapper database.Myocardial ischemic disease targets were searched by OMIM database.The potential therapeutic targets of Xinkeshu tablets against myocardial ischemia were analyzed.GO and KEGG enrichment analysis were conducted on core targets.The active ingredients were verified by zebrafish and cell model.qRT-PCR was used to detect the expression of key targets.Results Xinkeshu tablets could significantly alleviate ISO-induced pericardial edema and bradycardia.It also could increase sinus venous-bulb aortic(SV-BA)distance and improve the cell viability.The 30 potential active ingredients of Xinkeshu tables mainly acted on 30 core targets,including ALB,AKT1 and MAPK1,to regulate 627 GO items,including protein phosphorylation,negative regulation of apoptosis and positive regulation of PI3K signal transduction.KEGG results showed that 117 signaling pathways,including PI3K/Akt,FOXO and Ras,exerted anti-myocardial ischemia effect.Salvianolic acid A,lithospermic acid,rosmarinic acid,salvianolic acid D,salvianolic acid B,ginsenoside Rg2,hyperoside,3′-methoxypuerarin,3′-hydroxypuerarin and ginsenoside Rg1 could alleviate ISO-induced zebrafish myocardial ischemia and improve the cell viability.Xinkeshu tablets could upregulate the expression of genes such as ras and akt1,and downregulate the expression of genes such as mapk1 and mapk8.Conclusion The active ingredients,including salvianolic acid A in Xinkeshu tablets,exert anti-myocardial ischemia effects by targeting targets,such as AKT1,MAPK1,and regulating signaling pathways,such as PI3K/A

关 键 词：心可舒片 网络药理学 斑马鱼 H9C2 心肌缺血 盐酸异丙肾上腺素 

分 类 号：R-332[医药卫生] R282.71R319R329.411R542.2