基于网络药理学探究枸杞子-菊花药对治疗糖尿病视网膜病变的作用机制

Study on the Action Mechanism of Gouqizi-Juhua Pair on Diabetic Retinopathy Based on Network Pharmacology

目的:基于网络药理学探究枸杞子-菊花药对治疗糖尿病视网膜病变(diabetic retinopathy, DR)的干预机制。方法:利用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform, TCMSP)检索枸杞子、菊花的活性成分及相关靶点。利用Cytoscape 3.9.1绘制“药对-活性成分-作用靶点”网络图,并进行拓扑分析筛选枸杞子-菊花药对治疗DR的主要活性成分。利用GeneCards数据库、在线人类孟德尔遗传数据库及药物靶标数据库检索DR相关靶点。将活性成分相关靶点与疾病靶点导入Venny 2.1在线平台,绘制韦恩图,交集靶点即为枸杞子-菊花药对治疗DR的潜在靶点。将潜在靶点上传至STRING数据库构建PPI网络关系,导入Cytoscape 3.9.1软件进行可视化,PPI网络拓扑分析筛选枸杞子-菊花药对治疗DR的核心靶点。将核心靶点输入Metascape在线平台进行基因本体(gene ontology, GO)富集分析及京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)信号通路富集分析。将活性成分、核心靶点、信号通路导入Cytoscape 3.9.1软件构建“活性成分-作用靶点-信号通路-DR”网络图。结果:利用TCMSP数据库得到52种活性成分及247个相关靶点。“枸杞子菊花药对-活性成分-作用靶点”网络分析筛选出槲皮素、β-谷甾醇、山柰酚等为枸杞子-菊花药对治疗DR的主要活性成分。检索得到DR相关靶点2 347个,枸杞子-菊花药对治疗DR的潜在靶点133个。PPI网络分析筛选出蛋白激酶Bα(protein kinase Bα,AKT1)、半胱氨酸天冬氨酸蛋白酶3(caspase-3,CASP3)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)、前列腺素内过氧化物合酶2(prostaglandin-endoperoxide synthase 2,PTGS2)等为核心靶点。GO富集分析得到生物过程919个、细胞组分23个、分子功能47个。KEGG信号通路富集分析得到134条信号通路。“活性成分-作用靶点-信号通路-DR”网络分析发现枸杞子-菊花药对通过槲皮素、β-谷甾醇、山柰酚等活性成分作用于PTGS2、CASP3、AKT1等靶点调控糖尿病并发症中的AGE-RAGE信号通路、IL-17信号通路、TNF信号通路、PI3K-AKT信号通路、MAPK信号通路等进而改善DR。结论:枸杞子-菊花药对通过多成分、多靶点、多通路作用于DR。

Objective:To study the intervention mechanism of Gouqizi(Fructus Lyci)-Juhua(Flos Chrysanthemi)pair for the treatment of diabetic retinopathy(DR)based on network pharmacology.Methods:Traditional Chinese medicine systems pharmacology database and analysis(TCMSP)was used to search the active ingredients and related targets of Gouqizi and Juhua.Cytoscape 3.9.1 was used to construct the network diagram of"pair-active ingredients-target",and topological analysis was performed to screen the main active in-gredients of Gouqizi-Juhua pair in the treatment of DR.GeneCards database,online human Mendelian genetic database and drug target data were used to retrieve DR Related targets.Relevant targets of the active ingredients and disease targets were imported into Venny 2.1 online plaform to draw the Wayne diagram,and the intersection targets were potential targets of Gouqizi-Juhua pair for the treatment of DR.The potential targets were uploaded to STRING database to construct PPI network relationship,and imported into Cytoscape 3.9.1 software for visualization.PPI network topology analysis was performed to screen the core targets of Gouqizi-Juhua pair for DR treat-ment.The core targets were input into the Metascape online platform for gene ontology(GO)enrichment analysis and kyoto encyclope-dia of genes and genomes(KECG)signaling pathway enrichment analysis.The active ingredients,core targets,and signal pathways were imported into Cytoscape 3.9.1 software to construct the"active ingredient-target-signal pathway-DR"network diagram.Results:A total of 52 active ingredients and 433 related targets were obtained by TCMSP database.Quercetin,β-sitosterol and kaempferol were se-lected as the main active ingredients in the treatment of DR through the network analysis of"Gouqizi-Juhua pair-active ingredients-tar-get".A total of 2347 DR related targets and 133 potential targets of Gouqizi-Juhua pair were obtained.PPI network analysis screened protein kinase Bα(AKT1),caspase-3(CASP3),matrix metalloproteinase-9(MMP-9),prostaglandin-endoperoxidesynthase2(PTGS2)were the core targets.GO enrichment analysis obtained 919 biological processes,23 cellular components and 47 molecular functions.The enrichment analysis of KEGG signal pathways obtained 134 signal pathways.The"active ingredient-target-signal path-way-DR"network analysis showed that Gouqizi-Juhua pair regulated AGE-RAGE signaling pathway,IL-17 signaling pathway,TNF sig-naling pathway and PI3K-Akt signaling pathway MAPK signaling pathway in diabetic complications through the action of quercetin,β-sitosterol,kaempferol and other active ingredients on PTGS2,CASP3,AKT1 and other targets,which could improve DR.Conclusion:Gouqizi-Juhua pair acts on DR through multi-component,multi-target and multi-pathway.