To Investigate the Mechanism of Action of Artesunate in the Treatment of Fungal Keratitis Based on Network Pharmacology and Molecular Docking

目的：应用网络药理学方法探究青蒿琥酯(Artesunate, ART)治疗真菌性角膜炎(Fungal keratitis, FK)的作用机制。方法：通过SwissTagetPredictions数据库筛选青蒿琥酯的潜在作用靶点。采用GeneCards和OMIM数据库搜索与真菌性角膜炎相关的靶点。将数据合并去重后取药物靶点和疾病靶点的交集，获取青蒿琥酯治疗真菌性角膜炎的潜在作用靶点，应用Venny2.1平台绘制韦恩图。采用Cytoscape软件构建“青蒿琥酯–真菌性角膜炎–靶点”网络图。将交集靶点导入STRING数据库构建PPI蛋白互作网络并结合拓扑学分析筛选关键靶点。随后通过基因本体(GO)功能富集、京都基因与基因组百科全书(KEGG)通路富集分析对交集靶点进一步细化研究，最后使用Autodock软件对关键靶点与作用的活性化合物进行分子对接。结果：网络药理学，筛选青蒿琥酯作用靶点110个；筛选真菌性角膜炎靶点1809个；预测得到青蒿琥酯治疗真菌性角膜炎的潜在作用靶点共34个；蛋白互作网络分析发现，EGFR、STAT3、MMP9、SRC和STAT3可能是青蒿琥酯治疗真菌性角膜炎的核心靶点；经GO富集分析得到673个细胞生物学过程，主要涉及细胞对化学刺激的反应、细胞对氧化应激以及对非生物刺激的反应等生物学过程；KEGG通路富集分析获得79条相关信号通路，主要包括Relaxin、FoxO、TNF、HIF-1、Notch、Rap1等信号通路；分子对接结果显示，青蒿琥酯与EGFR的结合稳定性最好，与STAT3、MMP9、SRC和STAT3的结合比较稳定。结论：青蒿琥酯可能作用于EGFR、STAT3、MMP9、SRC和STAT3等靶点，通过Relaxin、FoxO、TNF、HIF-1、Notch、Rap1信号通路等发挥治疗真菌性角膜炎的作用。Objective: To investigate the mechanism of Fungal keratitis (FK) treated with artesunate (ART) by network pharmacology. Methods: Screening potential targets of ART using SwissTagetPredictions database. GeneCards and OMIM databases were used to search for targets associated with fungal keratitis. The intersection of drug targets and disease targets was obtained by combining the data and removing the data, and the potential targets of ART in the treatment of fungal keratitis were obtained. The Venny 2.1 platform was used to draw the Wayne map. The network diagram of “Artesunate-fungal keratitis-Target” was constructed by Cytoscape software. The intersection targets were imported into STRING database to construct PPI protein interaction network, and the key targets were screened by topological analysis. The intersection targets were further studied by gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Finally, Autodock software was used to perform molecular docking between key targets and active compounds. Results: 110 targets of ART were screened by network pharmacology. A total of 1809 targets of fungal keratitis were selected. A total of 34 potential targets of ART were predicted for the treatment of fungal keratitis. Protein interaction network analysis showed that EGFR, STAT3, MMP9, SRC and STAT3 may be the core targets of ART in the treatment of fungal keratitis. By GO enrichment analysis, 673 cell biological processes were obtained, which mainly involved cell response to chemical stimulation, cell response to oxidative stress and cell response to abiotic stimulation. KEGG pathway enrichment analysis obtained 79 related signaling pathways, including Relaxin, FoxO, TNF, HIF-1, Notch, Rap1 and other signaling pathways. Molecular docking results showed that ART had the best binding stability with EGFR, and was stable with STAT3, MMP9, SRC and STAT3. Conclusion: ART may act on EGFR, STAT3, MMP9, SRC and STAT3, and has therapeutic effects on fungal keratitis through Relaxin, FoxO, TNF, HIF-1, Notch and Rap1 signaling pathways.