基于网络药理学、分子对接和体外细胞实验研究白花蛇舌草总黄酮对食管鳞癌的作用机制

Mechanistic Investigation of Total Flavonoids from Hedyotis diffusa Willd. in Esophageal Squamous Cell Carcinoma via Network Pharmacology, Molecular Docking, and in Vitro Cell Experiments

摘要: 目的：借助网络药理学与分子对接方法，探究白花蛇舌草总黄酮抗食管鳞癌的作用机制，并通过体外细胞实验进行验证，为后续研究提供依据。方法：借助TCMSP获取白花蛇舌草的药物靶点，从GeneCards、TTD和OMIM数据库筛选食管鳞癌相关靶点，并利用Veen图获取其交集靶点；应用Cytoscape 3.10.2构建PPI网络，进行GO与KEGG富集分析；进一步将活性成分槲皮素(Quercetin)与核心靶点蛋白进行分子对接，并通过体外细胞实验验证作用效果。结果：共筛选出127个交集靶点，包括NFKB1、EGF、TP53、ERBB2等，确定Quercetin为有效成分。主要调控NFKB1、TP53及EGF信号通路发挥抗肿瘤作用。分子对接实验显示其与核心靶点结合良好；细胞实验表明白花蛇舌草总黄酮浓度提升会抑制食管鳞癌细胞存活。结论：白花蛇舌草总黄酮的抗肿瘤作用可能与其调控NFKB1与TP53等靶点相关。

Abstract: Objective: To investigate the mechanism of action of total flavonoids from Hedyotis diffusa Willd. (HF) against esophageal squamous cell carcinoma (ESCC) using network pharmacology and molecular docking methods, and to verify the effect through in vitro cell experiments, so as to provide a basis for subsequent research. Methods: The drug targets of Hedyotis diffusa were obtained from TCMSP. ESCC-related targets were screened from GeneCards, TTD, and OMIM databases, and the intersection targets were identified using a Venn diagram. Cytoscape 3.10.2 was used to construct a protein-protein interaction (PPI) network, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Furthermore, molecular docking was performed between the active component quercetin and core target proteins, and the effect was verified by in vitro cell experiments. Results: A total of 127 intersection targets were screened, including NFKB1, EGF, TP53, ERBB2, etc., and quercetin was identified as the effective component. It exerts anti-tumor effects mainly by regulating NFKB1, TP53, and EGF signaling pathways. Molecular docking experiments showed that quercetin binds well to core targets; cell experiments indicated that the inhibitory effect of HF on the viability of ESCC cells is concentration-dependent. Conclusion: The anti-tumor effect of total flavonoids from Hedyotis diffusa may be associated with their regulation of targets such as NFKB1 and TP53.