基于网络药理学和分子对接探讨绿原酸治疗 真菌性角膜炎的作用机制

Exploration on the Chlorogenic Acid in Treating Fungal Keratitis Based on Network Pharmacology and Molecular Docking

摘要: 目的：运用网络药理学和分子对接方法探讨绿原酸(Chlorogenic acid, CGA)治疗真菌性角膜炎(fungal keratitis, FK)的潜在作用机制。方法：通过SwissTargetPrediction等数据库检索绿原酸的潜在作用靶点，利用GeneCards、OMIM等数据库筛选真菌性角膜炎相关靶点，取交集后获得绿原酸治疗真菌性角膜炎的共同靶点，并绘制韦恩图。将共同靶点导入STRING数据库构建蛋白质相互作用(protein-protein interaction, PPI)网络，借助Cytoscape软件筛选核心靶点。进一步应用R软件进行基因本体论(Gene Ontology, GO)功能富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路富集分析，并采用AutoDock软件对核心靶点与绿原酸进行分子对接验证。结果：共筛得绿原酸与真菌性角膜炎的共同靶点93个。PPI网络分析显示，TNF、IL6、IL1B、AKT1、TP53、EGFR、MMP9、NFKB1、BCL2和ALB等靶点处于网络核心位置。GO富集分析结果提示，共同靶点主要涉及炎症反应调节、对外源刺激反应、细胞黏附正调控、MAPK级联正调控及氧化还原相关过程。KEGG富集分析显示，相关靶点主要富集于TNF signaling pathway、IL-17 signaling pathway、NOD-like receptor signaling pathway和HIF-1 signaling pathway等通路。分子对接结果表明，绿原酸与10个核心靶点均具有较好的结合活性，结合能均小于0 kcal/mol，其中与MMP9、AKT1、IL6和EGFR的结合活性相对较强。结论：本研究初步揭示了绿原酸治疗真菌性角膜炎具有多靶点、多通路协同作用的特点，其机制可能主要与调控炎症反应、减轻应激损伤及促进组织修复有关。本研究为进一步阐明绿原酸治疗真菌性角膜炎的分子机制及后续实验研究提供了理论依据。

Abstract: Objective: To investigate the potential mechanism of chlorogenic acid (CGA) in the treatment of fungal keratitis (FK) by using network pharmacology and molecular docking approaches. Methods: Potential targets of chlorogenic acid were retrieved from databases such as SwissTargetPrediction, while FK-related targets were collected from GeneCards, OMIM, and other databases. The intersection of drug targets and disease targets was identified as the common targets of CGA against FK, and a Venn diagram was constructed. These common targets were imported into the STRING database to establish a protein-protein interaction (PPI) network, and core targets were further screened using Cytoscape software. Subsequently, Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using R software. In addition, molecular docking between CGA and the core targets was conducted using AutoDock software for validation. Results: A total of 93 common targets between chlorogenic acid and fungal keratitis were identified. PPI network analysis showed that TNF, IL6, IL1B, AKT1, TP53, EGFR, MMP9, NFKB1, BCL2, and ALB were located at the core positions of the network. GO enrichment analysis indicated that the common targets were mainly involved in regulation of inflammatory response, response to exogenous stimuli, positive regulation of cell adhesion, positive regulation of the MAPK cascade, and oxidation-reduction-related processes. KEGG enrichment analysis revealed that the related targets were mainly enriched in the TNF signaling pathway, IL-17 signaling pathway, NOD-like receptor signaling pathway, and HIF-1 signaling pathway. Molecular docking results demonstrated that chlorogenic acid exhibited good binding activity with all 10 core targets, with binding energies all below 0 kcal/mol, among which the binding affinities with MMP9, AKT1, IL6, and EGFR were relatively stronger. Conclusion: This study preliminarily revealed that chlorogenic acid may exert therapeutic effects on fungal keratitis through a multi-target and multi-pathway synergistic mechanism. Its potential mechanism may be mainly related to the regulation of inflammatory responses, alleviation of stress-induced injury, and promotion of tissue repair. This study provides a theoretical basis for further elucidating the molecular mechanism of chlorogenic acid in the treatment of fungal keratitis and for subsequent experimental research.