基于网络药理学推测和分子生物学验证探讨半枝莲抗胰腺癌的活性成分及其作用机制

Active compounds and underlying mechanisms of Scutellariae Barbatae Herba against pancreatic cancer based on network pharmacology and molecular experimental validation

目的运用网络药理学预测和分子生物学验证探讨半枝莲抗胰腺癌的活性成分及其作用机制。方法通过中医药系统药理学数据库和分析平台(TCMSP)筛选半枝莲的活性成分和相关潜在靶点;应用毒性与基因比较数据库(CTD)、治疗靶点数据库(TTD)、药物基因组学知识库(PharmGKB)建立胰腺肿瘤靶点数据库;基于半枝莲潜在靶点与胰腺肿瘤靶点的匹配结果,构建PPI网络,拓扑分析筛选核心靶点;基于核心靶点的GO功能富集分析和KEGG通路富集分析建立半枝莲抗胰腺癌的信号通路图;采用Western blotting验证半枝莲中核心成分木犀草素对胰腺癌细胞PANC-1中P53、Bax和Bcl-2关键靶点的调控作用。结果共筛选出28种活性成分,包括槲皮素、木犀草素、汉黄芩素等;91个半枝莲作用靶点,经拓扑筛选获得24个核心靶点,涉及TP53、AKT1、JUN、VEGF等。GO功能富集分析显示,半枝莲作用靶点共涉及到73条生物学过程,KEGG通路富集筛选出18条信号通路参与胰腺癌的治疗,主要是通过NF-κB、p53、PI3K-Akt和VEGF信号通路来调节肿瘤炎性微环境、细胞周期阻滞、促细胞凋亡以及抗血管生成。分子生物学实验发现半枝莲中木犀草素可调节p53信号通路中关键靶点促进胰腺癌细胞的凋亡,证明了本研究中网络药理学分析结果的可靠性。结论初步证实了半枝莲具有多成分-多靶点-多途径协同作用的特点以及治疗胰腺肿瘤的潜在分子机制,为进一步探索其作用机制及临床应用奠定基础。

Objective To probe active components of Scutellariae Barbatae Herba(SBH)and its underlying complex mechanism in treating pancreatic cancer based on network pharmacology and molecular experimental validation.Methods Active compounds of SBH and potential targets of these compounds were screened via Traditional Chinese Medicine Systems Pharmacology(TCMSP)database.Then the pancreatic cancer target database was established by using Comparative Toxicogenomics Database(CTD),Therapeutic Target Database(TTD),and Pharmacogenomics Knowledgebase(PharmGKB).Based on the matching results between SBH potential targets and pancreatic cancer targets,a PPI network was structured and then the hub targets were screened by using topology analysis.Furthermore,DAVID bioinformatics was utilized for both Gene ontoloty(GO)functional enrichment analysis and KEGG pathway enrichment analysis.At last,Western blotting was used to validate the regulating function of luteolin on P53,Bax and Bcl-2 targets of PANC-1 cells.Results A total of 28 active ingredients including quercetin,luteolin and wogonin were screened out,and 24 hub targets of 91 targets including TP53,AKT1,JUN and VEGF were obtained.The results of DAVID enrichment analysis indicated that 73 cellular biological processes and 18 pathways were found to participate in the treatment of pancreatic cancer,mainly involving in regulating inflammatory microenvironment,cell cycle arrest,pro-apoptosis and anti-angiogenesis through NF-κB,p53,PI3K-AKT and VEGF signal pathways.Luteolin from SBH was validated to regulate key targets in the p53 signaling pathway to play a role in pro-apoptosis,which proved the reliability of results of network pharmacology.Conclusion This study confirmed the synergistic effect of multiple component-multiple target-multiple pathway of SBH on the treatment of pancreatic cancer,which offered a theory for its clinical application.