摘要
目的:利用网络药理学方法探讨癌痛消颗粒(ATXF)防治肝癌的物质基础和作用靶点,并结合大鼠移植性肝癌模型进行初步的验证。方法:通过TCMSP数据库收集癌痛消颗粒各中药活性成分及其潜在作用靶点,在GeneCards、OMIN、Drugbank、TTD数据库获取肝癌疾病靶点,取两者交集获得ATXF防治肝癌的潜在作用靶点。使用Cytoscape 3.8软件构建ATXF各成分与肝癌作用靶点的相互作用网络,并对其结果进行可视化分析。使用STRING平台分析ATXF防治肝癌靶点的互作网络,并运用CytoNCA分析各节点之间的拓扑关系,利用中位数筛选获得ATXF防治肝癌的核心靶点。通过Metascape数据分析平台,对获得的核心靶点进行GO、KEGG通路富集分析。采用大鼠腹水癌Walker-256细胞株建立SD大鼠移植性肝癌模型,经ATXF灌药干预15 d后取出瘤块。采用Western blot检测比较治疗组与对照组移植瘤内AKT、pAKT、p53、p-p53、ERK1/2以及p-ERK1/2的蛋白表达水平。结果:从ATXF中筛选得到257种活性成分,作用于294个靶点,与7993个肝癌疾病靶点相映射得到潜在作用靶点231个。进一步筛选获得AKT1、IL6、TP53、MAPK3、TNF、JUN、CASP3、MAPK1、MYC、PTGS2、MMP9等11个核心靶点,GO及KEGG分析结果显示以上核心靶点富集于HBV、TNF和癌症相关信号通路。大鼠移植瘤实验结果表明,ATXF可显著下调移植瘤内AKT、pAKT、pERK1/2的表达水平,同时显著上调p-p53蛋白表达水平,差异具有统计学意义(P<0.05)。结论:运用网络药理学方法分析发现癌痛消颗粒可作用于多条肿瘤信号通路,并运用大鼠移植瘤实验初步证实ATXF对于AKT、p53以及ERK1/2蛋白具有显著的调控作用,其机制可能是通过调控以上信号通路而发挥抗肝癌作用。
Objective:To investigate the mechanism of action and material basis of AiTongXiao granule in the treatment of hepatocellular carcinoma(HCC)based on network pharmacology and transplanted liver cancer rat model.Methods:TCMSP data⁃base was used to screen out effective components and its corresponding potential pharmaceutical targets,and databases including Gene Cards,OMIN,Drugbank and TTD were further used to collect HCC⁃related drug targets.The intersecting targets were ob⁃tained by mapping the drug and disease targets.The component⁃targets network was constructed and visualized by Cytoscape 3.8.2 software.Protein⁃protein interaction(PPI)network was built by STRING online platform,and the topological relationship and core targets were analyzed and screened by using CytoNCA software.In addition,Metascape database was used to perform gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis of the core tar⁃gets.At last,A transplanted liver cancer rat model was established by using Walker⁃256 cell line and treated by AiTongXiao gran⁃ule for 15 days.Western blot was used to further compare the expression levels of AKT,pAKT,p53,p⁃p53,ERK1/2 and ERK1/2 in the tumor between the treatment group and the control group.Results:A total of 257 active components were obtained from AiTongXiao granule,corresponding to 294 drug targets.Meanwhile,231 of the 7993 HCC disease targets were screened out between AiTongXiao granule drug and HCC disease targets.Eleven core targets including AKT1,IL6,TP53,MAPK3,TNF,JUN,CASP3,MAPK1,MYC,PTGS2 and MMP9 were further obtained by median screening.GO and KEGG analysis results showed that these core targets enriched to HBV,TNF and cancer related pathways.The rat transplanted liver cancer mod⁃el results indicated significant downregulation for AKT,p⁃AKT,pERK1/2,and significant upregulation of p⁃p53 after AiTongX⁃iao granule treatment(P<0.05).Conclusion:AiTongXiao granule could act to multiple cancer related pathways,and AKT,p53 and ERK1/2 were validated to be regulated by ATXF in rat model.The mechanism may be through the regulation of the above signaling pathways to exert anti⁃liver cancer effect.
作者
刘欢
刘显
金丽杰
刘莎莎
韦燕飞
LIU Huan;LIU Xian;JIN Li-jie;LIU Sha-sha;WEI Yan-fei(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530024,China;Guangxi Key Laboratory of Molecular Biology of Preventive Medicine of Traditional Chinese Medicine,Nanning 530024,China;Teaching Experiment Training Center of Guangxi University of Traditional Chinese Medicine,Nanning 530200,China;College of Basic Medicine of Guangxi University of Chinese Medicine,Nanning 530200,China)
出处
《海南医学院学报》
2023年第21期1632-1641,共10页
Journal of Hainan Medical University
基金
广西科技基地和人才专项(桂科AD20297013)
广西自然科学基金项目(2021GXNSFBA220036)
广西中医药大学第二批“岐黄工程”高层次人才团队培育项目(2021001)。
关键词
癌痛消颗粒
肝癌
大鼠移植瘤模型
网络药理学
信号转导
AiTongXiao granule
Hepatocellular carcinoma
Transplanted tumor rat model
Network pharmacology
Sig⁃nal transduction
