基于“成分-靶点-通路”的枸杞子调控年龄相关性黄斑变性作用机制研究

目的采用网络药理学方法探讨枸杞子改善年龄相关性黄斑变性(age-related macular degeneration,AMD)的活性成分及作用机制。方法以检索TCMSP数据库和查阅文献获得的枸杞子潜在活性成分为研究对象,通过Swiss Target Prediction平台、GeneCards数据库、OMIM数据库、DDT数据库和Drugbank数据库预测活性成分的潜在靶点;应用Metascape平台对潜在靶点进行基因本体(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)富集分析;运用String数据库和Cytoscape软件构建蛋白相互作用(protein-protein interaction,PPI)网络和"药物-成分-靶点-通路-疾病"网络;考察枸杞子不同提取部位对碘酸钠诱导的人视网膜色素上皮细胞ARPE-19活力的影响,以及对脂多糖诱导的小鼠小胶质细胞系BV2中炎症因子表达的影响。结果枸杞子中的88个活性成分通过调控血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、白细胞介素-8(interleukin-8,IL-8)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)等靶点,参与氧化应激、炎症反应等生物学过程,影响晚期糖基化终末产物(advanced glycation end products,AGE)-晚期糖基化终末产物受体(receptor for advanced glycation end products,RAGE)、IL-17、TNF等信号通路发挥改善AMD的作用。枸杞子95%乙醇提取部位能够显著提高碘酸钠诱导的ARPE-19细胞活力(P<0.001),并显著降低脂多糖诱导的BV2细胞中IL-6、IL-1β、TNF-αmRNA表达水平(P<0.05、0.01、0.001)。结论枸杞子中多种成分具有影响AMD相关靶点的潜在作用,枸杞子能够通过改善氧化损伤、抑制炎症反应治疗AMD,为后续深入研究枸杞子中不同活性成分调节AMD的作用机制提供依据。

Machansim of Lycii Fructus on regulating age-related macular degeneration based on “component-target-pathway”

Objective To explore the mechanism and active components of Lycii Fructus in treating age-related macular degeneration(AMD) based on network pharmacology and molecular experimental validation. Methods Active compounds of Lycii Fructus obtained from TCMSP database and literature investigation were selected as the research objects. Swiss Target Prediction platform, GeneCards, OMIM, DDT, and Drugbank database were used to screen and predict potential targets. Metascape platform was used for gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis of potential targets. The protein-protein interaction network model and "drug-ingredients-targets-pathways-disease" network were constructed by using String database and Cytoscape software. The effects of different extraction parts of Lycii Fructus on the activity of ARPE-19 of human retinal pigment epithelial cells induced by sodium iodate and the expression of inflammatory factors in mouse microglia cell line BV2 induced by lipopolysaccharide were investigated. Results A total of 88 active ingredients in Lycii Fructus might regulate vascular endothelial growth factor A(VEGFA), interleukin-8(IL-8), interleukin-6(IL-6), tumor necrosis factor(TNF), matrix metalloproteinase-9(MMP-9) and other targets. Potential targets mainly involve biological processes such as response to oxidative stress, regulation of inflammatory response, which played roles in the treatment of AMD by adjusting AGE-RAGE signaling pathway, IL-17 signaling pathway, and TNF signaling pathway. The results showed that the 95% ethanol extracts of Lycii Fructus significantly improved the survival rate of ARPE-19 cells damaged by sodium iodate(P < 0.001) and reduced the mRNA levels of IL-6, IL-1β, and TNF-α in BV2 cells induced by lipopolysaccharide(P < 0.05, 0.01, 0.001). Conclusion The results of network pharmacology showed that a variety of components in Lycii Fructus had potential effects on AMD related targets, and preliminarily proved that Lycii Fructus could treat AMD by improving oxidative damage and inhibiting inflammatory response, which can provide references for further research on the mechanism of AMD regulation by different active compounds in Lycii Fructus.