基于系统药理学探讨百合乌药汤对1型糖尿病并发肝损伤的保护作用

Protective Effect of Baihe Wuyao Decoction(百合乌药汤) in Treatment of Type 1 Diabetes Mellitus and Induced Hepatic Injury Based on Systems Pharmacology

目的:基于系统药理学及小鼠1型糖尿病(Type 1 diabetes mellitus,T1DM)模型探讨百合乌药汤对糖尿病并发肝损伤的保护作用。方法:通过中药系统药理数据库和分析平台(Traditional Chinese Medicines Systems Pharmacology,TCMSP)筛选百合乌药汤的活性成分;Swiss Target Prediction数据库结合查阅文献筛选出百合乌药汤的潜在活性成分和药物治疗靶点;通过OMIM、GeneCards数据库获取T1DM并发肝损伤作用靶点;使用Venn2.1获得药物-疾病共同靶点;采用Cytoscape 3.7.2软件构建“中药-成分-靶点-疾病”网络,借助Network Analyzer功能进行核心药物成分分析;使用STRING数据库进行蛋白相互作用分析(PPI);基于R软件使用Bioconductor生物信息软件包进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路分析;使用Cytoscape 3.7.2软件构建“中药-活性成分-靶点-信号通路”整合核心网络分析;采用AutoDock软件对百合乌药汤的潜在活性成分与关键靶标进行分子对接来验证结合性能;最后采用百合乌药汤干预T1DM并发肝损伤小鼠试验进行保肝作用和关键分子靶点的验证。结果:获得百合乌药汤包括植物甾醇、β-谷甾醇、等20个活性成分及589个靶点,与疾病的共同靶点有111个,包括蛋白激酶B1(AKT1)、丝裂原活化蛋白激酶(MAPK3)等;GO功能富集分析涉及生物过程包括蛋白丝氨酸/苏氨酸激酶活性的正向调节、MAP激酶活性的正向调节等;细胞组分表达过程中涉及膜筏、膜微区等;分子功能相关的过程中主要有跨膜受体蛋白激酶活性、跨膜受体蛋白酪氨酸激酶活性等;KEGG通路富集分析涉及糖尿病并发症中的AGE-RAGE、叉头盒子信号通路(FoxO)、PI3K-AKT等信号通路;采用T1DM并发肝损伤小鼠模型验证发现,百合乌药汤可改善T1DM并发肝损伤小鼠肝功能,上调磷酸化AKT蛋白的表达,改善胰岛素抵抗。结论:百合乌药汤可通过多成分、多靶点、多通路的作用特点治疗1型糖尿病。

Objective:To explore the protective effect of Baihe Wuyao Decoction(BWD)on type 1 diabetes mellitus(T1 DM)and induced hepatic injury in mice based on systems pharmacology.Methods:The active components of BWD were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).The potential active components and therapeutic targets of BWD were screened out from Swiss Target Prediction and literature reports.The related targets of T1 DM and induced hepatic injury were obtained through Online Mendelian Inheritance in Man(OMIM)and GeneCards.Venn 2.1 was used to obtain common drug-disease targets,and Cytoscape 3.7.2 was used to construct a“drug-component-target-disease”network.Network Analyzer was used to analyze core drug components,and STRING was used to analyze protein-protein interaction(PPI).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were carried out by Bioconductor based on R software.An integrated and core“drug-active component-target-pathway”network was constructed and analyzed by Cytoscape 3.7.2.AutoDock was applied for molecular dock of the potential active components of BWD to the key targets to verify the binding performance.Finally,the test of BWD intervention in the mouse model of T1 DM and induced hepatic injury was conducted to verify the hepatic protective effect and the key molecular targets.Results:Twenty active components and 589 targets from BWD were obtained.A total of 111 common targets related to T1 DM and induced hepatic injury were obtained,including protein kinase B1(AKT1)and mitogen-activated protein kinase(MAPK)3.In terms of GO enrichment analysis,biological processes involved positive regulation of serine/threonine protein kinase and MAPK activity.Cellular component expression involved membrane raft,membrane microdomains,etc.Molecular functions involved transmembrane receptor protein kinase activity,transmembrane receptor protein tyrosine kinase activity,etc.KEGG enrichment analysis showed signaling pathways involved included advanced glycation end-products and their receptors(AGE-RAGE),forkhead box O(FoxO),and phosphoinositide 3 kinase-protein kinase B(PI3 K-AKT)in diabetic complications.The integrated“drug-component-target-pathway”network with multi-component,multi-target,and multi-pathway characteristics was constructed.Molecular docking results showed that the core components in BWD had a good binding activity with the targets,which verified the accuracy of the prediction made by the network.The results of the animal experiment showed that BWD improved hepatic function,up-regulated the expression of phosphor-AKT/AKT(p-AKT/AKT),and improved the insulin resistance of the mouse model of T1 DM and induced hepatic injury.Conclusion:BWD can treat T1 DM in a multi-component,multi-target,and multi-pathway manner.