基于网络药理学与实验研究探讨黄芩苷对Hepa1⁃6肝癌细胞凋亡的影响  

Network pharmacology and experimental study⁃based research on effects of baicalin on apoptosis in Hepa1⁃6 hepatocellular carcinoma cells

作　　者：王玉洁 林晨 王一凤 祝峻峰 王见义 WANG Yujie;LIN Chen;WANG Yifeng;ZHU Junfeng;WANG Jianyi(Department of Hepatology,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院肝病科,上海200437

出　　处：《上海中医药杂志》2024年第7期31-39,共9页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市卫健委上海市中医药领军人才项目[ZY(2018-2020)-RCPY-1101]。

摘　　要：目的通过网络药理学方法并结合细胞实验,探讨黄芩提取物黄芩苷对肝癌细胞Hepa1-6恶性生物学行为的影响及其促进肝癌细胞凋亡的潜在作用机制。方法①通过中药系统药理学数据库与分析平台(TCMSP)等筛选黄芩苷的潜在化合物靶标;在人类基因综合数据库(Genecard)、人类孟德尔遗传数据库(OMIM)和药物治疗靶标数据库(TTD)中检索与肝癌的疾病靶点,将肝癌疾病靶点与黄芩苷化合物靶点整合,构建韦恩图及其蛋白质互作网络(PPI),以揭示黄芩苷治疗肝癌相关的关键靶点;利用Cytoscape软件选择交互基因,并利用David数据库、Metascape数据库分别进行基因本体(GO)功能富集和京都基因和基因组百科全书(KEGG)通路富集分析,阐明黄芩苷治疗肝癌的作用机制及相关通路。②通过细胞计数试剂盒(CCK-8)实验检测不同浓度的黄芩苷对Hepa1-6细胞活力的影响;Transwell细胞侵袭实验观察黄芩苷对Hepa1-6细胞侵袭能力的影响;流式细胞术检测不同浓度的黄芩苷对肝癌细胞Hepa1-6凋亡的影响;Western blot法检测白介素-6(IL-6)/信号转导与转录激活因子3(STAT3)信号通路相关蛋白及B淋巴细胞瘤-2蛋白(Bcl-2)、Bcl-2关联X蛋白(Bax)的表达情况。结果基于网络药理学共鉴定出黄芩苷作用靶点527个,肝癌疾病靶点1550个,双方交集靶点139个;综合网络药理学筛选结果选定Janus激酶(JAK)/STAT3信号通路作为研究通路。细胞实验表明,黄芩苷能够抑制肝癌细胞的增殖、侵袭和迁移等一系列恶性生物学行为,流式细胞术及Western blot结果表明黄芩苷可抑制肝癌细胞内IL-6/STAT3信号通路的激活,调控Bcl-2、Bax的表达而促进肝癌细胞的凋亡。结论网络药理学结合细胞实验研究发现黄芩苷是治疗肝癌的潜在有效药物,可以抑制肝癌细胞的增殖、侵袭、迁移活动,显著促进肝癌细胞凋亡,其作用机制与其下调IL-6/STAT3信号通路密切相关。Objective To investigate the effects of baicalin,an extract from Huangqin(Scutellariae Radix),on the malignant biological behaviors of Hepa1-6 hepatocellular carcinoma(HCC)cells and its potential mechanism in promoting apoptosis in HCC cells by combining network pharmacology and cellular experiments.Methods①The potential targets of baicalin were determined using the Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform(TCMSP).Subsequently,HCC related disease targets were retrieved from the Human Gene Database(GeneCards),the Online Mendelian Inheritance in Man(OMIM),and the Therapeutic Target Database(TTD).These targets were integrated with the baicalin compound targets to construct Venn diagrams and a protein-protein interaction(PPI)network to reveal key targets in baicalin treatment of HCC.Interaction genes were selected using Cytoscape software,and gene ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed using the Database for Annotation,Visualization and Integrated Discovery(DAVID)and Metascape database to elucidate the mechanism of action and related pathways in baicalin treatment of HCC.②The effects of different concentrations of baicalin on Hepa1-6 cell viability were assessed using Cell Counting Kit-8(CCK-8)assays.The impact on cell invasion capabilities was observed through Transwell cell invasion assays,and the effects of different concentrations of baicalin on the apoptosis of Hepa1-6 HCC cells were analyzed by flow cytometry.Western blot analysis was conducted to detect the expression of interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3)signaling pathway-related proteins,B-cell lymphoma 2(Bcl-2)and Bcl-2-associated X(Bax)proteins.Results Network pharmacology identified 527 baicalin targets and 1,550 HCC related disease targets,with 139 overlapping targets.The Janus kinase(JAK)/STAT3 signaling pathway was selected as the research pathway based on network pharmacology scr

关 键 词：肝癌 黄芩苷 细胞凋亡 细胞迁移 网络药理学 中药研究 作用机制 

分 类 号：R285[医药卫生—中药学]