基于网络药理学探讨党参、海藻“药对”治疗肝癌的作用机制及关键的靶点通路预测

目的:运用网络药理学方法筛选党参、海藻“药对”治疗肝癌主要化学成分，预测其作用靶点及通路。方法:借助中药系统药理分析数据库（TCMSP）、“中国知网”数据库检索党参、海藻“药对”的化学成分；TCMSP、PubChem、SEA以及Swiss Target Prediction数据库检索化学成分的靶点；通过TTD、人类孟德尔遗传数据库（OMIM）获取肝癌的相关作用靶点；运用Cytoscape3.8.2软件构建“党参、海藻-活性成分-靶点-肝癌”网络图；运用STRING数据库构建蛋白-蛋白相互作用（PPI）网络图；采用OmicsBean数据库进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）信号通路富集分析；采用四甲基偶氮唑盐微量酶反应比色法（MTT法）检测岩藻甾醇对人HepG2肝癌细胞增殖的抑制作用。结果:经数据库分析表明，党参的活性成分有20种、海藻6种，其中党参抗肝癌主要化合物是木犀草素、党参炔苷；海藻抗肝癌主要化合物是槲皮素、岩藻甾醇。党参、海藻“药对”化合物作用靶点共有964个，肝癌作用靶点有486个，化合物-肝癌共同作用靶点有32个，细胞表皮生长因子受体（EGFR）、蛋白激酶B（AKT1）、血管内皮生长因子A（VEGFA）与TP53为主要作用靶点。GO功能富集分析主要涉及细胞增殖、程序性死亡、凋亡负调控和催化等多个生物过程；胞质、细胞器和细胞核等结构；蛋白质二聚的分子功能。KEGG通路分析共得到信号通路190条，丝裂原活化蛋白激酶（MAPK）和磷脂酰肌醇3激酶（PI3K）-AKT为最主要的信号通路。MTT检测显示，与空白对照组（0 mmol/L）相比，岩藻甾醇各剂量组细胞存活率明显降低，差异均有显著性（均P<0.05）。结论:党参、海藻“药对”是多靶点、多通路治疗肝癌，为后续深入研究党参、海藻“药对”治疗肝癌的作用机制奠定基础，初步证明岩藻甾醇可以抑制人HepG2肝癌细胞的增殖。

Mechanism of and key target pathway prediction of Codonopsispilosula and Sargassum in the treatment of liver cancer based on network pharmacology

Objective: To screen the main chemical constituents of Codonopsispilosula and Sargassum "drug pair" in the treatment of liver cancer by the method of network pharmacology， and predict their action targets and pathways. Methods: The chemical constituents of Codonopsispilosula and Sargassum were searched by TCMSP and China knowledge Network database；the chemical composition targets were searched by TCMSP， PubChem， SEA and Swiss Target Prediction databases；the related targets of liver cancer were obtained by TTD and OMIM databases；the network map of "Codonopsispilosula， Sargassum-active ingredient-target-liver cancer" was constructed by Cytoscape3.8.2 software. Protein-protein interaction（PPI） network map was constructed by STRING database， gene ontology（GO） function enrichment analysis and Kyoto gene and genome encyclopedia（KEGG） signal pathway enrichment analysis were performed by Omics Bean database. The inhibitory effect of fucosterol on the proliferation of human HepG2 hepatoma cells was detected by tetrazolium salt microenzyme reaction colorimetry（MTT method）. Results: The database analysis showed that there were 20 kinds of active components in Codonopsispilosula and 6 species in Sargassum， among which the main anti-liver cancer compounds of Codonopsispilosula were luteolin and obetyolin， the main anti-liver cancer compounds of Sargassum were quercetin and fucosterol. There were 964 targets for drug pairs of Codonopsispilosula and Sargassum， 486 targets for liver cancer and 32 targets for compound-liver cancer interaction. Epidermal growth factor receptor（EGFR），protein kinases B（AKT1），Vascular endothelial cell growth factor A（VEGFA） and TP53 were the main targets. GO functional enrichment analysis mainly involved many biological processes such as cell proliferation， programmed death， negative regulation and catalysis of apoptosis， the structure of cytoplasm， organelle and nucleus， and the molecular function of protein dimerization. A total of 190 signal pathways were obtained by KEGG pathway analysis. MAPK and PI3K-AKT were the most important signal pathways. MTT detection showed that the cell survival rate of each dose of fucosterol was significantly lower than that of the blank control group（0 mmol/L）， and the difference was significant in a concentration-dependent manner. Conclusion: The "drug pair" of Codonopsispilosula and Sargassum is a multi-target and multi-pathway in the treatment of liver cancer， which lays a foundation for the further study of the mechanism of the "drug pair" of Codonopsispilosula and Sargassum in the treatment of liver cancer. It is preliminarily proved that fucosterol can inhibit the proliferation of human HepG2 hepatoma cells.