摘要
目的:从细胞实验和网络药理学探讨黄芪对肝癌的作用机制。方法:通过国家知识基础设施数据库(CNKI)、PubMed、中药系统药理学数据库与分析平台(TCMSP)、GeneCards数据库、在线人类孟德尔遗传数据库(OMIM)检索黄芪和肝癌的相关基因靶点及活性成分。通过STRING数据库和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)和黄芪-肝癌作用网络。以DAVID数据库进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)富集分析,并以CB-DOCK平台进行分子对接。细胞实验采用MTT法和蛋白质印迹法检测细胞增殖和蛋白表达。结果:筛选出黄芪19个有效成分,250个靶点。GO和KEGG富集分析得549个条目,145条信号通路,主要涉及调节DNA的转录,对细胞增殖和凋亡的调控等。作用通路主要涉及肿瘤相关的通路,如癌症通路、脂质和动脉粥样硬化、卡波西肉瘤、PI3K-AKT信号通路等。分子对接结果显示黄芪抗肝癌核心成分芒柄花黄素,毛蕊异黄酮,黄芪甲苷与PI3K-AKT信号通路蛋白有较好的结合力。网络药理学分析结果基础上,进一步细胞实验证实黄芪水提液能有效抑制HepG2细胞增殖,下调AKT1、PI3K p110,p-AKT1的蛋白表达水平。结论:黄芪能抑制PI3K-AKT信号通路,发挥对肝癌细胞的抑制作用,为其应用于肝癌临床治疗提供理论支持。
Objective:The mechanism of Astragalus on hepatocellular carcinoma was explored from the perspectives of cell experiment and network pharmacology.Methods:Relevant gene targets and active components of Astragalus and hepatocellular carcinoma were searched through CNKI,PubMed,TCMSP,GeneCards,and OMIM.The networks of protein-protein interaction(PPI)and Astragalus-hepatocellular carcinoma interaction were constructed by the STRING database and Cytoscape software.KEGG and GO enrichment analysis was performed with the DAVID database,and molecular docking was performed with the CB-DOCK platform.Cell experiments were performed by MTT assay and Western blot assay to detect cell proliferation and protein expression.Results:Nineteen active ingredients and 250 targets of Astragalus were screened out.GO and KEGG enrichment analysis obtained 549 entries and 145 signaling pathways,mainly involved in regulating DNA transcription,cell proliferation,and apoptosis.The pathway of action mainly involved tumor-related pathways,such as cancer pathway,lipid and atherosclerosis,Kaposi′s sarcoma infection,and PI3K-AKT signaling pathway.Molecular docking results showed that formononetin,isoflavones,and astragaloside IV,the core components of for anti-hepatocellular carcinoma of Astragalus,had good binding ability to proteins in the PI3K/AKT signaling pathway.Based on the results of network pharmacology analysis,further cell experiments confirmed that water extract of Astragalus can effectively inhibit the proliferation of HepG2 cells and down-regulate the protein expression levels of AKT1,PI3K p110,and p-AKT1.Conclusion:Astragalus can inhibit the PI3K/AKT signaling pathway,exert an inhibitory effect on HepG2 cells,and provide theoretical support for its application in the clinical treatment of hepatocellular carcinoma.
作者
庄耀隆
姚霏
侯文跃
张露蓉
宋卿
ZHUANG Yaolong;YAO Fei;HOU Wenyue;ZHANG Lurong;SONG Qing(Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Traditional Chinese Medicine,Suzhou 215009,China;Chinese Medicine Research and Development Department,Suzhou Wumen Medical School Research Institute,Suzhou 215009,China;Department of Oncology,Suzhou Hospital of Traditional Chinese Medicine,Suzhou 215009,China)
出处
《世界中医药》
CAS
2023年第23期3311-3316,共6页
World Chinese Medicine
基金
国家自然科学基金项目(82004136)
江苏省卫生健康委医学科项目(M2020004)。
关键词
黄芪
抗肝癌机制
PI3K-AKT信号通路
网络药理学
细胞实验
靶点
拓扑分析
分子对接
Astragalus
Anti-hepatocellular carcinoma mechanism
PI3K/AKT signaling pathway
Network pharmacology
Cell experiments
Target
Topology analysis
Molecular docking
