基于网络药理学和实验验证探究狼毒大戟醇提物治疗肝细胞癌的分子机制  

Mechanisms of Euphorbia fischeriana steud.alcohol extracts intreatment of HCC based on network pharmacology and experiment verification

作　　者：田辛晨 王浩辰 张一鸣 张家琪 张登甜 颜廷昊 赵静 蒋树龙 TIAN Xin-chen;WANG Hao-chen;ZHANG Yi-ming;ZHANG Jia-qi;ZHANG Deng-tian;YAN Ting-hao;ZHAO Jing;JIANG Shu-long(Cheeloo College of Medicine,Shandong University,Jinan 250000,China;Clinical Medical School,Jining Medical University,Jining Shandong 272000,China;Jining First People s Hospital,Jining Medical University,Jining Shandong 272000,China)

机构地区：[1]山东大学齐鲁医学院,山东济南250000 [2]济宁医学院临床医学院,山东济宁272000 [3]济宁医学院附属济宁市第一人民医院,山东济宁272000

出　　处：《中国药理学通报》2023年第7期1354-1361,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金项目(No81873249);山东省青年泰山学者资助计划(Notsqn20190922);山东省自然科学基金(NoZR2022MH319)。

摘　　要：目的 基于网络药理学和实验验证解析狼毒大戟醇提物治疗肝细胞癌作用机制。方法 通过TCMSP、Swiss ADME、Swiss Target Prediction数据库以及参考文献获得狼毒大戟醇提物活性成分和作用靶点;运用GeneCards和DisGeNET数据库筛选肝细胞癌相关基因;利用Venny平台、STRING平台和Cytoscape软件构建蛋白质-蛋白质互作网络图;使用DAVID数据库进行GO和KEGG富集分析。CCK-8、EdU和流式细胞术实验检测狼毒大戟醇提物对BEL-7402细胞增殖、凋亡的影响,Western blot分析狼毒大戟醇提物对JAK2/STAT3信号通路的作用,H22肝癌小鼠模型观察狼毒大戟醇提物的体内疗效。结果 共筛选获得狼毒大戟醇提物活性成分30个,潜在作用靶点567个,肝细胞癌疾病靶点916个,药物-疾病交集靶点115个;KEGG富集分析得到主要信号通路JAK2/STAT3;体外实验显示狼毒大戟醇提物能够抑制BEL-7402细胞增殖,促进其凋亡,并呈浓度依赖性的抑制JAK2/STAT3信号通路;此外,狼毒大戟醇提物单独或联合索拉非尼显著抑制H22肝癌小鼠肿瘤的生长。结论 狼毒大戟醇提物具有抗肝癌治疗作用,其分子机制可能与调控JAK2/STAT3信号通路有关。Aim To investigate the molecular mechanisms of alcohol extracts of Euphorbia fischeriana steud.against hepatocellular carcinoma(HCC)through a combination of network pharmacology analysis and experimental validation.Methods The active ingredients and targets of alcohol extracts of Euphorbia fischeriana steud.were determined through TCMSP,Swiss ADME,Swiss Target Prediction database and references.The databases DisGeNET and GeneCards were employed to screen potential HCC-related genes.Venny platform,STRING platform and Cytoscape software were applied to construct active ingredient-target-disease and protein-protein interaction(PPI)network maps.Gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed using the DAVID database.To assess the effects of Euphorbia fischeriana steud.alcohol extracts on BEL-7402 cells,the proliferation and apoptosis were detected by CCK-8,EdU and flow cytometry assays,and the related protein levels of JAK2/STAT3 pathway were analyzed by Western blot.Additionally,H22 hepatocellular carcinoma mouse model was used to evaluate the in vivo efficacy of Euphorbia fischeriana steud.alcohol extracts.Results A total of 916 HCC targeted genes,30 active ingredients containing the related 567 potential targeted genes,and 115 intersection targets of disease and compounds were obtained.KEGG enrichment analysis identified JAK2/STAT3 signaling as a critical pathway.In vitro experiments showed the alcohol extracts of Euphorbia fischeriana steud.could inhibit proliferation,promote apoptosis and suppress JAK2/STAT3 signaling pathway in a dose-dependent manner in BEL-7402 cells.In addition,the alcohol extracts of Euphorbia fischeriana steud.,either alone or in combination with sorafenib,dramatically blocked tumor growth in in vivo tests.Conclusions Euphorbia fischeriana steud.alcohol extracts have anti-cancer effects in HCC,and the molecular mechanisms may be connected to the regulation of JAK2/STAT3 signaling pathway.

关 键 词：肝细胞癌 狼毒大戟醇提物 药物靶点 分子对接 JAK2/STAT3信号通路 网络药理学 

分 类 号：R282.71[医药卫生—中药学] R319[医药卫生—中医学] R329.28R345.57R735.7