探讨葛根素滴眼液对慢性闭角型青光眼的作用机制

Based on the network pharmacology and molecular docking technology to investigate the potential mechanical effect of puerarin eye drops on chronic angle-closure glaucoma

目的基于网络药理学及分子对接技术探讨葛根素滴眼液的主要活性成分葛根素对慢性闭角型青光眼(CACG)的潜在作用机制和关键靶点。方法利用葛根素的结构信息通过PharmMapper数据库和SwissTarget数据库,结合中药系统药理学数据库和分析平台(TCMSP)检索葛根素的治疗靶点,同时通过GeneCards、OMIM和DisGeNET数据库获取CACG相关靶点,葛根素治疗靶点和CACG相关靶点取交集,得到葛根素滴眼液治疗CACG的潜在靶点。在String平台上进行蛋白互作分析,利用Cytoscape_v3.9.1软件对结果进行可视化及计算,得到葛根素滴眼液治疗CACG的关键靶点。使用David平台对葛根素滴眼液治疗CACG的潜在靶点进行基因本体(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析葛根素滴眼液治疗CACG的潜在作用机制。采用分子对接验证葛根素滴眼液与关键靶点的结合作用。结果经筛选显示,葛根素滴眼液有效成分的葛根素潜在治疗靶点共402个,CACG的靶点共得到1506个,葛根素抗CACG的潜在作用靶点共68个,其中关键靶点共8个。GO富集分析表明这些靶点主要与细胞凋亡过程、稳态维持、对缺氧的反应等生物过程有关,KEGG富集分析显示,主要通路为内分泌抵抗、松弛素信号通路、HIF-1信号通路、AGE-RAGE通路等。关键靶点分别为血管内皮生长因子A、TP53结合蛋白1、胱天蛋白酶3、肿瘤坏死因子、RAC-α丝氨酸/苏氨酸蛋白激酶、缺氧诱导因子1、连环蛋白β1、金属基质蛋白酶9。分子对接结果显示葛根素与除连环蛋白β1外的7个靶点之间均具有较好的结合活性。结论葛根素滴眼液可能作用于VEGFA、TP53、CASP3、TNF、AKT1、HIF1A、MMP9等靶点,通过调控细胞凋亡、PI3K-Akt信号通路、HIF-1信号通路、松弛素信号通路等信号通路起到治疗CACG作用。

Objective Based on the network pharmacology and molecular docking technology to investigate the potential mechanical effect and key target of puerarin,the major active constituent in puerarin eye drops,on chronic angle-closure glaucoma(CACG).Methods The therapeutic targets of puerarin were searched by PharmMapper database,SwissTarget database,TCM Systematic pharmacology database and Analysis Platform(TCMSP)by.At the same time CACG related targets were accessed through GeneCards,OMIM and DisGeNET.The potential target of treating CACG with puerarin eye drops was obtained by intersection of puerarin treatment target and CACG related target.Protein interaction analysis was carried out on the String platform,Cytoscape_v3.9.1 software was used to visualize and calculate the results,and the key targets of the treatment of CACG with puerarin eye drops were obtained.The David platform was used to analyze the potential targets’pathway enrichment of the gene ontology(GO)function and the Kyoto Encyclopedia of Genes and Genomes(KEGG)to understand potential mechanism of puerarin eye drops in the treatment of CACG.Results The screening results showed that there were 402 potential therapeutic targets of puerarin,1506 targets of CACG and 68 potential anti-CACG targets of puerarin,including 8 key targets.GO enrichment analysis showed that these targets were mainly related to biological processes such as apoptosis,homeostasis maintenance,and response to hypoxia.KEGG enrichment analysis showed that the main pathways were endocrine resistance,relaxin signaling pathway,HIF-1 signaling pathway,and AGE-RAGE pathway.The key targets were vascular endothelial growth factor A,TP53 binding protein 1,caspase 3,tumor necrosis factor,RAC-αserine/threonine protein kinase,hypoxia-inducing factor 1,cateninβ1,and metallomatrix proteinase 9.Molecular docking results showed that puerarin had good binding activity with 7 targets except cateninβ1.Conclusion Puerarin eye drops may act on VEGFA,TP53,CASP3,TNF,AKT1,HIF1A,MMP9 and other targets,and play a role in the treatment of CACG by regulating apoptosis,PI3K-Akt signaling pathway,HIF-1 signaling pathway,relaxin signaling pathway and other signaling pathways.