基于网络药理学、分子对接与实验验证揭示薯蓣皂苷元治疗动脉粥样硬化的作用机制  

Mechanism of diosgenin in treatment of atherosclerosis based on network pharmacology,molecular docking and experimental validation

作　　者：王丹 王肖龙[1,2] WAND Dan;WANG Xiao-long(Branch of National Clinical Research Center for Chinese Medicine Cardiology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Cardiovascular Research Institute of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,国家中医心血管病临床医学研究中心分中心,上海201203 [2]上海中医药大学附属曙光医院心血管病研究所,上海201203

出　　处：《中草药》2022年第24期7783-7794,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81573647);国家自然科学基金面上项目(82074222);上海市中医临床重点实验室(14DZ2273200);上海市科委引导类项目(19401934300);上海市卫健委中医药事业发展三年行动计划(ZY[2018-2020]-CCCX-2003-07);上海市临床重点专科项目(shslczdzk05301)。

摘　　要：目的通过网络药理学、分子对接结合体外验证的方法,探究薯蓣皂苷元抗动脉粥样硬化(atherosclerosis,AS)的潜在靶点及作用机制。方法通过SwissTargetPrediction、Pharmmapper、CheMBL数据库挖掘薯蓣皂苷元作用靶点,然后通过OMIM、Drugbank、Genecards、DisGeNET数据库预测与AS相关的靶点,取两者共同靶点,构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络;通过Cytoscape 3.7.2软件进行可视化,筛选核心靶点并进行基因本体(gene ontology,GO)功能和京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析;使用AutodockTools、PyMOL和Discovery Studio软件进行分子对接以验证薯蓣皂苷元和靶点的相关性。通过构建体外泡沫细胞模型,考察薯蓣皂苷元对泡沫细胞脂质代谢、促炎细胞因子水平及关键靶点表达的影响。结果共获得588个薯蓣皂苷元靶点和5489个与AS相关的靶点,获得275个薯蓣皂苷元与AS的交叉靶点,通过PPI网络筛选获得Src、信号传导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)等52个关键靶点。GO功能富集分析表明,这些靶点主要与蛋白质磷酸化、细胞对脂质的反应,细胞迁移的正向调节、炎症反应等过程相关。KEGG通路富集分析发现,薯蓣皂苷元抗AS与脂质和动脉粥样硬化通路、核因子-κB(nuclear factor-κB,NF-κB)信号通路等有关。分子对接表明,薯蓣皂苷元与Src、STAT3具有良好的结合亲和力。体外实验表明,薯蓣皂苷元显著抑制THP-1巨噬细胞脂质堆积和促炎细胞因子水平(P<0.05、0.01、0.001),显著下调Src和STAT3的磷酸化水平(P<0.05、0.01、0.001)。结论薯蓣皂苷元能够通过调节多个靶点和通路发挥抗AS作用,其作用机制可能与调节Src/STAT3通路从而改善脂质代谢和炎症有关。Objective To explore the potential targets and mechanism of diosgenin against atherosclerosis(AS)through network pharmacology,molecular docking and in vitro verification.Methods The targets of diosgenin were mined by SwissTargetPrediction,Pharmmapper and CheMBL databases,and then the targets related to AS were predicted by OMIM,Drugbank,Genecards and DisGeNET databases,and protein-protein interaction(PPI)network was constructed by taking the common targets of them.Cytoscape 3.7.2 software was used to visualize,then core targets were screened out and performed gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis.AutodockTools,PyMOL and Discovery Studio software were used for molecular docking to verify the correlation between diosgenin and targets.The effects of diosgenin on lipid metabolism,levels of pro-inflammatory cytokines and expressions of key targets in foam cells were investigated by constructing foam cell model in vitro.Results A total of 588 targets of diosgenin and 5489 targets related to AS were obtained,and 275 cross targets between diosgenin and AS were obtained.Through PPI network screening,52 key targets such as Src,signal transducer and activator of transcription 3(STAT3)were obtained.GO function enrichment analysis showed that these targets were mainly related to the phosphorylation of protein,reaction of cells to lipids,positive regulation of cell migration,inflammatory reaction and other processes.KEGG pathway enrichment analysis showed that the effect of diosgenin on AS was related to lipid and atherosclerosis pathway,nuclear factor-κB(NF-κB)signaling pathway,etc.Molecular docking showed that diosgenin had good binding affinity with Src and STAT3.In vitro experiments showed that diosgenin significantly inhibited THP-1 macrophage lipid accumulation and pro-inflammatory cytokine levels(P<0.05,0.01,0.001),significantly up-regulated Src and STAT3 phosphorylation levels(P<0.05,0.01,0.001).Conclusion Diosgenin plays an anti-AS role by regulating multiple

关 键 词：薯蓣皂苷元 动脉粥样硬化 网络药理学 分子对接 SRC 信号传导及转录激活蛋白3 

分 类 号：R285.5[医药卫生—中药学]