基于网络药理学和细胞验证探讨郁金-水红花子抗肝癌机制

Mechanism of Radix Curcumae and Fructus Polygoni Orientalis against liver cancer based on network pharmacology and cell verification

目的基于网络药理学预测并通过细胞实验验证探究郁金—水红花子对肝癌的作用机制。方法通过TCMSP筛选郁金—水红花子的药效活性成分,整理并校正其相应作用靶点。通过Ctdbase数据库收集肝癌相关基因靶点,运用Venny筛选郁金—水红花子治疗肝癌的关键靶点,利用STRING数据库对关键靶点进行蛋白相互作用(PPI)网络分析及GO和KEGG通路富集分析,采用Cytoscape构建成分—靶点—疾病网络。检测不同浓度郁金—水红花子提取物对HepG2细胞存活率的影响,采用Western blot检测HepG2细胞Akt、p-Akt、PI3K及p-PI3K蛋白表达。结果筛选郁金—水红花子活性成分28个,作用靶点278个,肝癌靶点35317个。PPI网络拓扑分析获得45个核心基因,通过GO及KEGG富集分析得到郁金—水红花子干预肝癌的关键信号通路。体外实验表明,郁金—水红花子对HepG2细胞增殖具有显著抑制作用,可下调Akt、p-Akt、PI3K及p-PI3K蛋白表达。结论郁金—水红花子对HepG2细胞增殖具有显著抑制作用,调控PI3K/Akt信号通路抑制肝癌细胞增殖可能是其部分作用机制。

Objective Exploring the mechanism of action of Radix Curcumae and Fructus Polygoni Orientalis on liver cancer based on network pharmacology prediction and cell experiment validation.Methods Screening the pharmacological active ingredients of Radix Curcumae and Fructus Polygoni Orientalis through TCMSP,organizing and correcting their corresponding target points of action.The liver cancer related gene targets through the Ctdbase database were searched.Using Venny to screen key targets for the treatment of liver cancer with Radix Curcumae and Fructus Polygoni Orientalis.Using STRING database for protein interaction(PPI)network analysis and GO and KEGG pathway enrichment analysis of key targets.Constructing a Component Target Disease Network Using Cytoscape.Detecting the effect of different concentrations of Radix Curcumae and Fructus Polygoni Orientalis extract on the survival rate of HepG2 cells.Using Western blot for detection of Akt,p-Akt,PI3K,and p-PI3K protein expression in HepG2 cells.Results Screening 28 active ingredients from Radix Curcumae and Fructus Polygoni Orientalis,with 547 targets for action and 35317 targets for liver cancer.45 core genes obtained through PPI network topology analysis.Key signaling pathways of Radix Curcumae and Fructus Polygoni Orientalis intervention in liver cancer identified through GO and KEGG enrichment analysis.In vitro experiments,it was showed that Radix Curcumae and Fructus Polygoni Orientalis has a significant inhibitory effect on HepG2 cell proliferation,downregulating the expression of Akt,p-Akt,PI3K,and p-PI3K proteins.Conclusion Radix Curcumae and Fructus Polygoni Orientalis has a significant inhibitory effect on liver cancer,and regulating the PI3K/Akt signaling pathway to inhibit liver cancer cell proliferation may be a partial mechanism of its action.