摘要
基于网络药理学及分子对接探讨黄芪抗肝癌的活性成分与分子作用机制。通过TCMSP数据库获取黄芪活性成分,Swiss Target Prediction预测成分靶点,采用Genecards数据库与OMIM数据库搜集肝癌靶点,Venny相映射黄芪抗肝癌的作用靶点,String数据库结合Cytoscape 3.7.2软件绘制肝癌靶点的蛋白相互作用网络(PPI)及“黄芪-成分-通路-肝癌”相互作用网络,DAVID数据库对核心靶点基因功能富集和通路富集分析。Surflex-Dock软件对黄芪关键成分与核心靶点进行分子对接验证。MTT法检测槲皮素、毛蕊异黄酮、山奈酚、芒柄花素、异鼠李素及华良姜素对肝癌细胞(HepG2)的影响。RT-qPCR法验证华良姜素对TP53、MAPK1、AKT1、IL6、MAPK8与VEGFA基因表达相对水平。本研究筛选出20个黄芪活性成分,涉及202个作用靶点及其100条KEGG信号通路,GO分析为487条生物功能。网络药理学分析黄芪可能是通过TP53、MAPK1、AKT1、IL6、MAPK8与VEGFA等关键靶点起到抗肝癌作用。分子对接表明靶点与成分有一定的结合性。MTT表明华良姜素对肝癌细胞(HepG2)的抑制作用较强于其他五个成分。RT-qPCR验证不同浓度的华良姜素对6个基因的表达量均为上调趋势,与KEGG通路分析所涉及基因一致。本研究初步探讨黄芪治疗肝癌具有多靶点、多通路的潜在作用机制,为后续验证黄芪抗肝癌的分子机制提供了依据。
Based on network pharmacology and molecular docking to explore the active ingredients and molecular mechanism of Astragalus Radix anti-liver cancer.Acquire the active components of Astragalus Radix from TCMSP database,predict component targets by Swiss Target Prediction,use Genecards database and OMIM database to collect liver cancer targets,Venny phase maps Astragalus Radix anti-liver cancer targets,String database combined with Cytoscape 3.7.2 software to draw liver cancer targets the protein interaction network(PPI)and the"Astragalus Radix-component-pathway-liver cancer"interaction network,the DAVID database analyzes the function enrichment and pathway enrichment of core target genes.Surflex-Dock software verifies the molecular docking between key components of Astragalus Radix and core targets.MTT method was used to detect the effects of quercetin,calycosin,kaempferol,formononetin,isorhamnetin and jaranol on liver cancer cells(HepG2).The RT-qPCR method verified the relative levels of Jaranol on the expression of TP53,MAPK1,AKT1,IL6,MAPK8 and VEGFA genes.In this study,20 active components of Astragalus Radix were screened,involving 202 targets and 100 KEGG signal pathways,and GO analysis showed 487 biological functions.Network pharmacology analysis Astragalus Radix may play an anti-liver cancer effect through key targets such as TP53,MAPK1,AKT1,IL6,MAPK8 and VEGFA.Molecular docking shows that the target and the component have a certain degree of binding.MTT showed that Jaranol has a stronger inhibitory effect on liver cancer cells(HepG2)than the other five components.RT-qPCR verified that the expression levels of the six genes of Jaranol at different concentrations were all up-regulated,which was consistent with the genes involved in the KEGG pathway analysis.This study preliminarily explored the potential mechanism of Astragalus Radix with multiple targets and multiple pathways in the treatment of liver cancer,and provided a basis for subsequent verification of the molecular mechanism of Astragalus Radix against liver cancer.
作者
姚红
任晋宏
侯宇芯
王禹璇
薛慧清
李青山
YAO Hong;REN Jin-hong;HOU Yu-xin;WANG Yu-xuan;XUE Hui-qing;LI Qing-shan(Shanxi University of Traditional Chinese Medicine,Shanxi Key Laboratory of Innovative Drugs for the Treatment of Serious Diseases Basing on the Chronic Inflammation,Taiyuan 030619,China)
出处
《天然产物研究与开发》
CAS
CSCD
2021年第6期1020-1031,共12页
Natural Product Research and Development
基金
国家国际合作专项(2013DFA30700)
山西省应用基础研究项目(201801D221437)。
