基于网络药理学及实验验证探究当归补血汤治疗糖尿病性视网膜病变的作用机制

Study on the mechanism of Danggui Buxue decoction in the treatment of diabetic retinopathy based on network pharmacology and experiment

本研究使用网络药理学、分子对接结合实验验证探究当归补血汤在糖尿病视网膜病变(Diabetic retinopathy,DR)中的作用机制。从TCMSP、TCMID、Swiss Target Prediction数据库筛选当归补血汤活性成分及其靶点,通过Gene Cards、Dis Ge NET收集DR靶点,利用Venny 2.1获得二者交集靶点。使用Cytoscape构建“疾病-药物-成分-靶点”、蛋白互作(PPI)网络;DAVID数据库对关键靶点进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析;Auto Dock Vina对主要活性成分与关键靶点进行分子对接验证。最后,通过动物实验验证网络药理学结果。收集到当归补血汤与DR共同靶点84个。GO分析显示220个条目;KEGG富集分析得到113条信号通路。分子对接结果显示关键靶点与槲皮素、豆甾醇等有效成分均具有较好的对接活性。动物实验表明当归补血汤可改善DR大鼠视网膜组织形态,降低血糖,增加胰岛素分泌,降低DR大鼠视网膜组织VEGF、TNF-α蛋白表达水平、提高PI3K、Akt1表达水平(P<0.05,P<0.01)。当归补血汤治疗DR具有多活性成分、多靶点、多通路的特点,可通过调节TNF-α、VEGFA、PI3K、AKT等核心靶点,影响PI3K/AKT、TNF等信号通路发挥对DR的治疗作用。

In the present study,we investigated the mechanism of action of Danggui Buxue decoction(DBD)in diabetic retinopathy(DR)using network pharmacology and molecular docking combined with experimental validation.GeneCards and DisGeNET were used to collect DR targets.The intersection of the two targets was obtained using Venny 2.1.Cytoscape was used to construct"disease-drug-component-target"and protein-protein interaction(PPI)networks.DAVID database was used to enrich key targets with Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).AutoDockVina was used to perform molecular docking of primary active ingredients with key targets.Molecular docking validation was performed with key targets by AutoDockVina.Finally,the network pharmacological results were validated by animal experiments.A total of 84 common targets of DBD and DR were collected,220 entries were revealed by GO analysis,and 113 signaling pathways were obtained by KEGG enrichment analysis.Molecular docking results showed that the key targets had suitable docking activities with quercetin,dousterol and other active ingredients.Animal experiments showed that DBD could improve retinal tissue morphology,lower blood glucose,increase insulin secretion,reduce VEGF and TNF-αprotein expression levels and increase PI3K and Akt1 expression levels in the retinal tissues of DR rats(P<0.05,P<0.01).The treatment of DR by DBD had the characteristics of multiple active ingredients,multiple targets and multiple pathways,which could exert therapeutic effects on DR by regulating the core targets of TNF-α,VEGFA,PI3K,and AKT and affecting the signaling pathways of PI3K/AKT and TNF.