摘要
目的:利用超高效液相色谱-四极杆飞行时间串联质谱(UPLC-Q-TOF-MS)和网络药理学探究黄芪-附子治疗心力衰竭的作用机制和物质基础。方法:运用UPLC-Q-TOF-MS技术分析黄芪-附子溶液中的化学成分,通过PubChem数据库筛选活性成分及作用靶点,利用比较毒性基因组学数据库(CTD)、在线人类孟德尔遗传数据库(OMIM)和GeneCard数据库检索心力衰竭疾病靶点,通过Venn分析获得共有靶点。利用STRING数据库分析靶点蛋白质-蛋白质相互作用(PPI)关系,利用Metascape数据库进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,采用SYBYL-X2.1.1软件对关键靶点和活性成分进行分子对接分析。通过大鼠心力衰竭模型对关键靶标进行实验验证。结果:黄芪-附子溶液中共鉴定出202个化学成分,经预测其中64个活性成分作用于183个靶点治疗心力衰竭,重要的活性成分有咖啡酸、L-精氨酸、鹰嘴豆芽素A、腺嘌呤、烟酸、反式阿魏酸、对香豆酸、核黄素、毛蕊异黄酮等,主要的作用靶点有白细胞介素(IL)-6、胱天蛋白酶(Caspase)-3、血管内皮生长因子受体A(VEGFA)、蛋白激酶B1(Akt1)、肿瘤坏死因子(TNF)、IL-1B、基质金属蛋白酶(MMP)-9等,主要涉及缺氧诱导因子-1(HIF-1)信号通路、TNF信号通路、磷脂酰肌醇3-激酶(PI3K)/Akt信号通路、Toll样受体信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、叉头框蛋白O(FoxO)信号通路等。分子对接结果显示黄芪-附子溶液中的活性成分与HIF-1信号通路中关键蛋白HIF-1α、VEGFA、Akt1、Caspase-3、IL-6有较好的结合能力。动物实验结果显示,黄芪-附子溶液能显著改善心力衰竭大鼠血流动力学指标,降低血清心钠肽(ANP)、脑钠素(BNP)和IL-6含量,改善心肌组织病理学变化,保护心肌细胞线粒体形态,下调心力衰竭大鼠心肌组织中HIF-1信号通路关键蛋白HIF-1α、VEGFA、磷酸化(p)-Akt的表达,降低Caspase-3的活化。结论:黄芪-附子通过多成分、多靶点、多途径治疗心力衰竭,实验验证说明其可通过改善心肌组织病理学变化、调节HIF-1信号通路发挥治疗心力衰竭的作用,为后续的药效物质基础研究提供了重要参考。
Objective:To explore the mechanism of Astragali Radix-Aconiti Lateralis Radix Praeparata in the treatment of heart failure and substance basis based on ultra-performance liquid chromatographyquadrupole-time of flight-mass spectrometer(UPLC-Q-TOF-MS)and network pharmacology.Method:The chemical components of Astragali Radix-Aconiti Lateralis Radix Praeparata solution was analyzed by UPLC-QTOF-MS,and the active components and targets were screened out by the PubChem database.The targets related to heart failure disease were retrieved from Comparative Toxicogenomics Database(CTD),Online Mendelian Inheritance in Man(OMIM),and GeneCard databases,and the common targets were obtained by Venn analysis.The target protein-protein interactions(PPI)were analyzed using the STRING database.Gene ontology(GO)functional enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed using the Metascape database,and molecular docking verification of key targets and active components was performed using SYBYL-X 2.1.1.Experimental validation of key targets was carried out using the rat model of heart failure.Result:There were 202 chemical components identified in Astragali Radix-Aconiti Lateralis Radix Praeparata solution,of which 64 active components were predicted to act on 183 targets for the treatment of heart failure.The important active components were caffeic acid,Larginine,biochanin A,adenine,nicotinic acid,trans-ferulic acid,p-coumaric acid,riboflavin,calycosin,etc.The main targets were interleukin(IL)-6,cysteine aspartic acid specific protease(Caspase)-3,vascular endothelial growth factor A(VEGFA),protein kinase B1(Akt1),tumor necrosis factor(TNF),IL-1B,matrix metallopeptidase(MMP)-9,etc.The main signaling pathways involved hypoxia-inducible factor(HIF)-1signaling pathway,TNF signaling pathway,phosphatidylinositol 3-kinase(PI3K)/Akt signaling pathway,Toll-like receptor signaling pathway,mitogen-activated protein kinase(MAPK)signaling pathway,forkhead box O(FoxO)signaling pathway,etc.The molecular docking results showed that the active components in Astragali Radix-Aconiti Lateralis Radix Praeparata solution had a good binding ability with HIF-1α,VEGFA,Akt1,Caspase-3,and IL-6,which were the key proteins in the HIF-1 signaling pathway.Animal experiments showed that Astragali Radix-Aconiti Lateralis Radix Praeparata solution significantly improved the hemodynamic indexes,reduced the serum atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),and IL-6 levels,improved the myocardial histopathological changes,protected the mitochondrial morphology of cardiomyocytes,down-regulated the expression of HIF-1α,VEGFA and phosphorylation(p)-Akt,and reduced the activation of Caspase-3 in the myocardial tissue of rats with heart failure.Conclusion:Astragali RadixAconiti Lateralis Radix Praeparata treats heart failure in a multi-component,multi-target,and multi-pathway manner.The experimental validation indicates that it treats heart failure by improving myocardial histopathological changes and regulating HIF-1 signaling pathway,which provides references for the subsequent pharmacodynamic substance research.
作者
钟勰
张瑶
李军
毛涵
陈向云
李尧锋
ZHONG Xie;ZHANG Yao;LI Jun;MAO Han;CHEN Xiangyun;LI Yaofeng(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine(TCM),Guiyang 550025,China;Resource Institute for Chinese&Ethnic Materia Medica of Guizhou University of TCM,Guiyang 550025,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第3期70-80,共11页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金地区科学基金项目(81960864,81860873)
贵州省高层次创新型人才“千层次”人才项目(贵中医[ZQ2018005])
贵州省科技计划项目(黔科合基础[2016]1401)
贵州中医药大学学术新苗培养及创新探索专项项目(黔科合平台人才[2017]5735号-06)。
关键词
心力衰竭
黄芪
附子
网络药理学
缺氧诱导因子-1(HIF-1)信号通路
heart failure
Astragali Radix
Aconiti Lateralis Radix Praeparata
network pharmacology
hypoxia-inducible factor-1(HIF-1)signaling pathway
