基于网络药理学和分子对接探讨瑞香素治疗 真菌性角膜炎的作用机制

Mechanism of Daphnetin in the Treatment of Fungal Keratitis Based on Network Pharmacology and Molecular Docking

摘要: 目的：应用网络药理学方法研究瑞香素(Daphnetin)治疗真菌性角膜炎(Fungal keratitis, FK)的潜在作用机制。方法：从SwissTargetPrediction数据库中查询瑞香素的潜在作用靶点，应用OMIM和GeneCards数据库检索真菌性角膜炎的相关治疗靶点。将数据库数据合并去重后，通过Venny 2.1.0平台绘制韦恩图获取交集靶点。利用STRING平台构建蛋白质相互作用(PPI)网络，将结果导入Cytoscape 3.10.3软件进行可视化分析及核心靶点筛选。使用R软件对交集靶点进行基因本体论(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。最后使用Autodock vina对核心靶点与瑞香素进行分子对接验证。结果：共获得瑞香素潜在作用靶点58个，真菌性角膜炎相关靶点1114个，瑞香素治疗真菌性角膜炎的潜在交集靶点15个。PPI网络包含15个节点和50条边，经拓扑分析筛选出SRC、EGFR、AKT1、PTK2、BRAF等核心靶点。GO功能富集分析显示靶点主要涉及蛋白磷酸化正调控、肽基–酪氨酸磷酸化等生物过程，以及膜筏、细胞前缘等细胞组分和蛋白酪氨酸激酶活性等分子功能。KEGG通路富集分析显示靶点显著富集于黏着斑通路、EGFR耐药通路、ErbB信号通路、PI3K-Akt信号通路、MAPK信号通路等。分子对接结果显示瑞香素与SRC (−6.4 kcal/mol)、EGFR (−7.3 kcal/mol)、AKT1 (−5.9 kcal/mol)、PTK2 (−6.7 kcal/mol)、BRAF (−6.1 kcal/mol)均具有较好的结合活性。结论：本研究预测了瑞香素治疗真菌性角膜炎的15个潜在靶点，并初步探讨了其可能通过作用于SRC、EGFR、AKT1、PTK2、BRAF等核心靶点，调控黏着斑通路、PI3K-Akt通路和MAPK通路，影响免疫细胞迁移、炎症反应和氧化应激等过程，从而发挥治疗作用。这为理解瑞香素治疗真菌性角膜炎的作用机制提供了科学依据和参考。

Abstract: Objective: To investigate the potential molecular mechanism of Daphnetin in the treatment of fungal keratitis (FK) using a network pharmacology approach. Methods: Potential targets of Daphnetin were retrieved from the SwissTargetPrediction database, while disease-associated targets of fungal keratitis were collected from OMIM and GeneCards databases. After merging and removing duplicates, overlapping targets were identified and visualized using the Venny 2.1.0 platform. A protein-protein interaction (PPI) network was constructed via the STRING database and further analyzed using Cytoscape 3.10.3 for visualization and screening of core targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using R software. Finally, molecular docking was conducted via AutoDock Vina to verify the binding affinity between Daphnetin and the identified core targets. Results: A total of 58 potential targets for Daphnetin and 1114 targets related to fungal keratitis were identified, resulting in 15 overlapping targets for the treatment of FK. The PPI network contained 15 nodes and 50 edges. Through topological analysis, core targets such as SRC, EGFR, AKT1, PTK2, and BRAF were identified. GO enrichment analysis indicated that these targets are primarily involved in biological processes, such as the positive regulation of protein phosphorylation and peptidyl-tyrosine phosphorylation; cellular components including membrane rafts and the cell leading edge; and molecular functions such as protein tyrosine kinase activity. KEGG analysis revealed that the targets were significantly enriched in the Focal adhesion, EGFR tyrosine kinase inhibitor resistance, ErbB signaling pathway, PI3K-Akt signaling pathway, and MAPK signaling pathway. Molecular docking results showed that Daphnetin exhibited favorable binding activities with SRC (−6.4 kcal/mol), EGFR (−7.3 kcal/mol), AKT1 (−5.9 kcal/mol), PTK2 (−6.7 kcal/mol), and BRAF (−6.1 kcal/mol). Conclusion: This study predicted 15 potential targets for Daphnetin in treating fungal keratitis. The findings suggest that Daphnetin may exert its therapeutic effects by acting on core targets, such as SRC, EGFR, AKT1, PTK2, and BRAF, thereby modulating pathways like Focal adhesion, PI3K-Akt, and MAPK. These interactions likely influence immune cell migration, inflammatory responses, and oxidative stress, providing a scientific basis and reference for understanding the therapeutic mechanism of Daphnetin in fungal keratitis.