基于代谢组学与网络药理学的白花蛇舌草抑制胃癌 作用机制研究

Study of the Pharmacological Basis and Mechanism of Hedyotis diffusa Willd. in Suppressing Gastric Cancer based on NMR Metabolomics combined Network Pharmacology approach

关键词:  白花蛇舌草  胃癌  NMR代谢组学  网络药理学  作用机制  有效成分

Key words:  hedyotis diffusa willd.  gastric cancer  NMR metabolomics  network pharmacology.

摘要:

摘要：目的 该研究通过联合代谢组学和网络药理学探讨白花蛇舌草（Hedyotis diffusa Willd., HDW）抗胃癌的活性成分及机制。方法 利用TCMSP数据库并结合已发表文献收集HDW的化学成分，同时基于Swiss Target Prediction数据库获取其靶基因，并使用GeneCards数据库获取胃癌靶点基因；通过Cytoscape软件绘制HDW -胃癌共有靶点网络，在STRING数据库构建共有靶点的蛋白-蛋白相互作用（PPI）网络，在DAVID数据库进行基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）通路富集分析，筛选HDW对胃癌的活性成分及靶点。同时，使用AGS胃癌细胞进行体外实验，测试HDW对AGS细胞的增殖、迁移及侵袭的抑制作用，确定HDW抗胃癌的半数有效浓度（IC50）；基于核磁共振代谢组学方法分析细胞代谢物，探讨HDW的作用机制。结果 结果显示HDW能明显抑制AGS细胞划痕伤口的愈合率及侵袭能力，其对AGS细胞的IC50为4.495 mg/mL。通过细胞代谢物代谢组学分析，筛选到15个潜在生物标记物与HDW的抑制作用有关，主要涉及到与能量代谢有关的6条代谢通路。此外，网络药理学结果显示，HDW与胃癌共有靶点175个，结合PPI交互网络筛选到5个核心靶点。结论 HDW可能通过反式咖啡酸、反式阿魏酸、山奈酚、异麦芽酮糖、槲皮素和香豆酸等成分，TNF、EGFR、STAT3、MMP9和PTGS2等靶点，调节能量代谢、ROS产生与炎症反应等通路治疗胃癌。

Abstract:

ABSTRACT: OBJECTIVE To investigate the active components and mechanism of Hedyotis diffusa Willd. (HDW) against gastric cancer by metabolomics and network pharmacology research methods. METHODS The TCMSP database and published documents were used to obtain the chemical compounds of HDW, while SwissTarget Prediction database was used to obtain the target genes of HDW and target genes of gastric cancer were obtained using GeneCards database. Then, to screen the active compounds and key targets of HDW, common targets network of HDW and gastric cancer was visualized by Cytoscape, and the protein-protein interaction (PPI) network of common targets was constructed via STRING database. Gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were analyzed in the DAVID database. Meanwhile, the in vitro experiments were conducted using human gastric adenocarcinoma cells (AGS cell line) to test the inhibitory effects of HDW on the proliferation, migration and invasion of AGS cells, and to determine the half effective concentration (IC50) of HDW against gastric cancer. In addition, metabolomics analysis of cellular metabolites was performed using nuclear magnetic resonance (NMR) technology to explore the potential mechanism of HDW. RESULTS The IC50 of HDW on AGS cells was 4.495 mg/mL, which significantly inhibited the wound healing rate in the scratch test and inhibited the invasive ability of AGS cells, with the better effect than that of cisplatin. 15 potential biomarkers were screened that associated with the inhibition of HDW and involved in 6 metabolic pathways related to energy metabolism. In addition, the results of network pharmacology showed that there were 175 common targets between HDW and gastric cancer, and 5 core targets were recognized combining the result of PPI interaction network. CONCLUSION Trans-caffeic acid, trans-ferulic acid, kaempferol, isomaltulose, quercetin and coumaric acid may be the potential active components of HDW in against gastric cancer, acting through the core targets including TNF, EGFR, STAT3, MMP9 and PTGS2, and regulating energy metabolism, ROS production and inflammation response.