基于UPLC-Q-TOF-MS^(E)联用网络药理学、分子对接分析肾衰泄浊汤干预CKD合并As的化学成分及作用机制

Chemical composition and mechanism of action of Shenshuai Xiezhuo Decoction based upon UPLC-QTOF-MS^(E)plus network pharmacology and molecular docking for treating chronic kidney disease with atherosclerosis

目的:检测肾衰泄浊汤主要活性成分,并预测肾衰泄浊汤治疗慢性肾脏病(chronic kidney disease,CKD)合并动脉粥样硬化(atherosclero sis,As)关键靶标与作用机制。方法:运用高效液相色谱-四极杆飞行时间串联质谱法(UPLC-Q-TOFMSE)与中药系统药理学数据库与分析平台/中药分子机理的生物信息学分析工具(TCMSP/BATMAN-TCM)数据库分析肾衰泄浊汤主要活性成分与靶标名称、数量等信息。通过在线人类孟德尔遗传(OMIM)、基因疾病关联数据库(DisGeNET)、人类基因综合数据库(GeneCards)数据库筛选疾病靶标,与药物靶标取交集,构建“中药-活性成分-作用靶点”网络,应用蛋白质/基因互作数据库(String)构建蛋白互作网络(PPI),基因本体论(Go)生物学功能注释和京都基因与基因组百科全书(KEGG)分析潜在作用机制;分子对接初步验证网络药理学结果。结果:UPLC-Q-TOF-MSE鉴定出33个活性成分,数据库检索获得75个活性成分,药物成分靶标349个,疾病靶标4241个,得到249个交集靶标。获得槲皮素、山柰酚、β-谷甾醇、木犀草素、异鼠李素等核心活性成分与蛋白激酶1(Akt1)、丝裂原活化蛋白激酶3(MAPK3)、信号转导和转录激活因子3(STAT3)等核心靶标;GO分析共获得1292条目;KEGG示潜在靶标主要富集在PI3K-AKT通路、脂质与动脉粥样硬化途径、糖尿病并发症相关AGE-RAGE信号通路等。分子对接表明核心成分与关键靶标结合良好。结论:肾衰泄浊汤活性成分可能通过干预MAPK3、Akt1、STAT3等靶标,调控炎症、氧化应激、脂质代谢等机制治疗CKD合并As,其中PI3K/Akt可能是重要的调节通路。

OBJECTIVE To detect the main active ingredients of Shenshuai Xiezhuo Decoction(SSXZ)and predict the mechanism of action and key targets of SSXZ for chronic kidney disease(CKD)with atherosclerosis(AS).METHODS Highperformance liquid chromatography quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS^(E))and traditional Chinese medicine system pharmacology database and analysis platform/bioinformatics analysis tool for traditional Chinese medicine molecular mechanism(TCMSP/BATMAN-TCM)were employed for examining the main active ingredients and target information of SSXZ.Disease targets were screened through online human Mendelian genetics(OMIM),gene disease association database(DisGeNET)and human gene comprehensive database(GeneCards),intersecting with drug targets for obtaining potential targets,constructing a"traditional Chinese medicine active ingredient target"network,using protein/gene interaction database(String)database for constructing a protein interaction network(PPI).Gene Ontology(Go)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were utilized for elucidating potential mechanisms of action.Finally molecular docking was verified between the core components of traditional Chinese medicines and key targets.RESULTS UPLC-Q-TOF-MSE yielded 33 active ingredients and 75 active ingredients from database search.There were 349 drug ingredient targets,4,241 disease targets and 249 intersection targets.The core active ingredients such as quercetin,kaempferol,β-sitosterol,lignocerol,isorhamnetin and the core targets of protein kinase 1(AKT1),mitogen-activated protein kinase 3(MAPK3)and signal transducer and activator of transcription 3(STAT3)were obtained;A total of 1,292 entries were obtained from GO analysis.KEGG indicated that potential targets were clustered largely in PI3K-AKT pathway,lipid and atherosclerosis pathway and AGE-RAGE signaling pathway correlated with diabetic complications.Molecular docking revealed that the core components conjugated well with key targets.CONCLUSION The active components of SSXZ may regulate inflammation,oxidative stress,lipid metabolism and other mechanisms through interfering with MAPK3,AKT1,STAT3 and other targets for CKD As.And PI3K/Akt may be an important regulatory pathway.