基于网络药理学和细胞验证探讨甘草抗肺癌机制

Mechanism of Glycyrrhizae Radix et Rhizoma against Lung Cancer Based on Network Pharmacology and Cell Verification

目的基于网络药理学探究甘草对肺癌的作用机制,并进行细胞实验验证。方法通过TCMSP筛选甘草药效活性成分,整理并校正其相应作用靶点,通过GeneCards、OMIM、DrugBank数据库收集肺癌相关基因靶点,筛选甘草治疗肺癌的关键靶点,利用STRING、DAVID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构建靶点-化合物网络。检测不同浓度甘草提取物对A549细胞存活率的影响,采用Western blot检测A549细胞Bax、Bcl-2、HIF-1α及GLUT1蛋白表达。结果筛选甘草活性成分88个,作用靶点213个,肺癌靶点23458个,匹配映射得到关键靶点209个。通过疾病-药物活性成分-作用靶点网络和PPI网络分析筛选出与甘草关联程度较大的靶点43个。GO富集分析得到生物过程1202条、细胞组分46条、分子功能64条。KEGG通路富集分析得到甘草治疗肺癌的通路226条,主要包括癌症相关通路、HIF-1信号通路、动脉粥样硬化、小细胞肺癌、幽门螺杆菌感染的上皮细胞信号转导等。体外实验表明,甘草提取物对A549肺癌细胞增殖具有显著抑制作用,可上调Bax蛋白表达,下调Bcl-2、HIF-1α及GLUT1蛋白表达。结论本研究通过网络药理学分析得到甘草治疗肺癌的活性成分、作用靶点及信号通路,可为甘草的抗癌活性及作用机制研究奠定基础。

Objective To explore the mechanism of Glycyrrhizae Radix et Rhizoma against lung cancer based on network pharmacology,and verify by cell experiment.Methods The active components of Glycyrrhizae Radix et Rhizoma were screened through TCMSP,and the corresponding action targets were corrected.The gene targets related to lung cancer were collected by GeneCards,OMIM and Drugbank databases,and the key targets of Glycyrrhizae Radix et Rhizoma in the treatment of lung cancer were screened.The key targets were transmitted to STRING database and DAVID database for PPI network analysis,GO enrichment analysis and KEGG pathway analysis,and constructed by Cytoscape.The effects of different concentrations of Glycyrrhizae Radix et Rhizoma extract on the survival rate of A549 cells were detected.The expressions of Bax,Bcl-2,HIF-1αand GLUT1 in A549 cells were detected by Western blot.Results Totally 88 active components of Glycyrrhizae Radix et Rhizoma,213 action targets and 23458 lung cancer targets were screened,and 209 key targets were obtained by matching mapping.Through the analysis of disease-drug active components-action target network and PPI network,43 targets with high correlation with Glycyrrhizae Radix et Rhizoma were selected.GO enrichment analysis revealed 1202 biological processes,46 cell components,and 64 molecular functions.KEGG pathway analysis showed that there were 226 main pathways of Glycyrrhizae Radix et Rhizoma in the treatment of lung cancer,including pathways in cancer,HIF-1 signaling pathway,atherosclerosis,small cell lung cancer,Helicobacter pylori infection and so on.In vitro experiments showed that Glycyrrhizae Radix et Rhizoma extract could significantly inhibit the proliferation of A549 lung cancer cells,up-regulate the expression of Bax protein and down-regulate the expression of Bcl-2,HIF-1αand GLUT1 protein.Conclusion Based on network pharmacology,this study preliminarily discusses the active components,action targets and signaling pathways of Glycyrrhizae Radix et Rhizoma in the treatment of lung cancer,and lays the foundation for the study of anticancer activity and mechanism of Glycyrrhizae Radix et Rhizoma.