摘要
目的基于网络药理学、分子对接和体外实验验证探索知母皂苷AⅢ(TIA)治疗胶质母细胞瘤(GBM)的潜在靶点及其信号通路。方法利用Pharmmapper数据库预测TIA的作用靶点,通过GeneCards数据库获得GBM相关的疾病靶点;通过STRING数据库得到交集靶点的蛋白-蛋白相互作用(PPI)网络图,并使用Cytoscape 3.6.0软件进行筛选和分析;交集靶点通过DAVID数据库进行基因本体(GO)功能注释分析和京都基因和基因组百科全书(KEGG)通路富集分析,采用分子对接技术考察TIA与关键靶点的结合能力。最后,采用GBM细胞U-87MG、U251进行体外细胞实验,运用细胞计数试剂盒(CCK-8)实验检测细胞存活率、Transwell迁移测定实验检测细胞迁移能力,采用Western blot实验、实时荧光定量逆转录聚合酶链式反应(RT-qPCR)实验及药物亲和反应的靶点稳定性(DARTS)实验等验证网络药理学分析结果。结果网络药理学预测结果显示,TIA作用靶点与GBM疾病靶点存在65个潜在共用靶点;GO分析结果主要集中在炎症应答、细胞增殖的正向调节等过程;KEGG富集分析显示,TIA通过免疫球蛋白G Fcγ受体(FcγR)介导的吞噬作用、谷胱甘肽代谢等信号通路发挥作用;分子对接分析结果显示,TIA与基质金属蛋白酶-2(MMP-2)的结合潜能最强。体外细胞实验结果显示,TIA呈选择性和剂量依赖性抑制人GBM细胞U-87MG和U251的体外增殖和迁移,并通过靶向亲和MMP-2,抑制其表达而发挥作用。结论TIA可能通过靶向结合MMP-2并抑制其表达而抑制GBM细胞的体外增殖和迁移。
Objective To explore the potential targets and signaling pathways of timosaponin AⅢ(TIA)in treating glioblastoma(GBM)based on network pharmacology,molecular docking,and in vitro experimental verification.Methods The Pharmmapper database was utilized to predict the targets of TIA,while disease-related targets of GBM were obtained from the GeneCards database.The protein-protein interaction(PPI)network of intersecting targets was acquired through the STRING database,and Cytoscape 3.6.0 software was used to screen and analysis.The intersecting targets were analyzed using the DAVID database for gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.Molecular docking technology was used to investigate the binding ability of TIA to key targets.Finally,GSM cells U-87MG and U251 were used for in vitro cell experiments.Cell Counting Kit-8(CCK-8),Transwell migration assay were used to measure cell survival rate and migration ability.Western blot,Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-qPCR),and drug affinity responsive target stability(DARTS)experiments were used to verify network pharmacological analysis results.Results The network pharmacological prediction results showed that there were 65 potential shared targets between TIA targets and GBM disease targets.The results of GO analysis mainly focused on the inflammatory response and the positive regulation of cell proliferation.KEGG enrichment analysis demonstrated that TIA exerts its effects through immunoglobulin G Fcγreceptor(FcγR)mediated phagocytosis,glutathione metabolism,and other signaling pathways.Molecular docking analysis indicated that TIA exhibited the strongest binding potential with metalloproteinase-2(MMP-2).The in vitro experimental results showed that TIA selectively and dosedependently inhibited the proliferation and migration of human GBM cells U-87MG and U251 in vitro,and works by inhibiting the expression of MMP⁃2 via targeting its affinity.Conclusi
作者
廖雅芳
孙辉
袁春露
苑艳丽
赵述达
章丹丹
LIAO Yafang;SUN Hui;YUAN Chunlu;YUAN Yanli;ZHAO Shuda;ZHANG Dandan(Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Oncology,Shanghai Lung Hospital Affiliated to Tongji University,Shanghai 200433,China)
出处
《上海中医药杂志》
CSCD
2024年第6期73-81,共9页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(81773946,82374116,81573673,81001666)。
关键词
胶质母细胞瘤
知母皂苷AⅢ
基质金属蛋白酶
网络药理学
分子对接
中药研究
glioblastoma
timosaponin AⅢ
matrix metalloproteinase
network pharmacology
molecular docking
traditional Chinese herbal medicine research
