基于生物信息技术探究骨碎补-金银花治疗糖尿病足的作用机制

Research on the mechanism of Rhizoma Drynariae-Lonicera japonica in treating diabetic foot based on bioinformation technology

目的:通过网络药理学及动物实验研究骨碎补-金银花治疗糖尿病足的作用机制。方法:通过TCMSP网站获取骨碎补-金银花的药物成分及作用靶点,将靶点通过uniprot统一基因名称;通过GeneCards查询糖尿病足的疾病靶点,利用venny 2.1获取交集靶点。将交集靶点导入STRING进行分析并使用Cytoscape构建PPI网络图。将交集靶点导入Metascape分析GO富集和KEGG富集。并通过Cytoscape软件构建"成分-靶点-通路"网络图。依据Degree分别筛选PPI中前4位基因靶点,并通过动物实验验证骨碎补-金银花干预糖尿病足大鼠动物模型后血清中靶蛋白表达差异。结果:骨碎补-金银花32个有效成分调节148个疾病靶点治疗糖尿病足,其中甾醇、豆甾醇、木樨草素、山奈酚等是核心成分,JUN、TP53、蛋白激酶B(Akt1)、丝裂原活化蛋白激酶(MAPK)3是至关重要的靶点。基因功能注释分析结果显示,与氧化应激、膜筏、膜微区、质膜筏、激酶结合、蛋白激酶结合等相关。KEGG分析结果提示骨碎补-金银花主要参与MAPK、PI3K/Akt等信号通路。动物实验证实糖尿病足大鼠血清JUN、TP53、Akt1、MAPK3表达增加。结论:骨碎补-金银花主要通过调节MAPK、PI3K/Akt等信号通路的JUN、TP53、Akt1、MAPK3等疾病靶点,干预酶的活性、炎症反应等生物学过程进而治疗糖尿病足。

Objective:To study the mechanism of Rhizoma Drynariae-Lonicera japonica on diabetic foot through network pharmacology and animal experiment. Methods:The drug components and targets of Rhizoma Drynariae-Lonicera japonica were obtained from TCMSP website and the targets were converted into gene names through UniPROt database. GeneCards was used to query disease targets of diabetic foot,and venny 2.1 was used to obtain intersection targets. The intersection targets were imported into STRING for protein interaction analysis and PPI network graph was constructed using Cytoscape. The intersection targets were imported into Metascape to analyze GO enrichment and KEGG enrichment. The "component-target-pathway" network was constructed by Cytoscape software. The first four PPI gene targets were screened according to Degree,and the difference of target protein expression in serum after Rhizoma Drynariae-Lonicera japonica intervention in diabetic foot rat animal model was verified by animal experiments. Results:The screening of 32 active components of beta-sitosterol,stigmasterol,luteolin and kaempferol could regulate multiple pathways and 148 disease targets in the treatment of diabetic foot,of which beta-sitosterol,stigmasterol,luteolin and kaempferol were the core components,and JUN,TP53,Akt1 and MAPK3were the most important targets. Gene function annotation analysis showed that it was related to,oxidative stress,membrane rafts,membrane microregions,plasma membrane rafts,kinase binding,protein kinase binding,etc. The results of KEGG analysis suggested that Rhizoma Drynariae-Lonicera japonica were mainly involved in MAPK and PI3K/Akt signaling pathways. Animal experiments confirmed that the expressions of JUN,TP53,Akt1 and MAPK3 in serum of diabetic foot rats were increased. Conclusion:The treatment of Rhizoma Drynariae-Lonicera japonicafor diabetic foot is mainly through regulating JUN,TP53,Akt1,MAPK3 and other disease targets of MAPK,PI3K/Akt signaling pathways,and intervening the biological processes of enzyme activity and inflammatory response.