摘要
目的阐释古代经典名方“保元汤”的药效物质基础并探讨其分子水平的作用机制,发掘保元汤更多潜在的临床应用优势。方法采用UPLC-QTOF-MS/MS技术,选用Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水(A)-乙腈(B)为流动相系统进行梯度洗脱,高分辨飞行时间质谱电喷雾电离源(ESI),正、负离子扫描模式,检测鉴定保元汤的化学成分组成,并分析保元汤大鼠多次多天ig给药后含药血浆中的原型化学成分及代谢产物,进一步运用网络药理学方法,利用TCMSP数据库和药物信息数据库等分析保元汤入血原型成分的靶向作用,并揭示其治疗潜能。结果根据精确相对分子质量数据和多级质谱碎片离子,结合对照品、数据库以及文献报道,共鉴定出保元汤中133个化学成分,并且确定大鼠ig保元汤后,35个成分能够经口吸收入血达到血浆稳态的药物浓度。保元汤入血原型成分“成分-靶点”网络图、核心靶基因基因本体(gene ontology,GO)功能富集分析、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析、疾病预测分析表明,保元汤可通过人参皂苷Rb1、丹皮酚、刺芒柄花素、刺甘草查耳酮、大豆异黄酮、甘草查耳酮A、6-姜辣素、水杨酸等8个关键成分作用于丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、肿瘤蛋白p53(tumor protein p53,TP53)、信号传导及转录激活蛋白(signal transducer and activator of transcription)等核心靶点,通过糖基化终末产物(advanced glycation end products,AGE)-晚期糖基化终末产物受体(receptor for advanced glycation end-products,RAGE)、表皮生长因子受体(epidermal growth factor receptor,EGFR)、非受体酪氨酸激酶(janus kinase,JAK)-信号转导子和转录激活子(signal transducer and activator of transcription,STAT)等信号通路等途径发挥药效作用,提示保元汤除治疗慢性心力衰竭外,还对阿尔茨海默症、再灌注损伤、绝经后骨质疏松症等具有潜在治疗优势。保元汤35个入血原型成分中含有多个同类成分的衍生物,且其中既有单一成分对接多个靶点、又有多个类似物对接同一靶点的情况,由此佐证了“中药(复方)多成分多靶点协同作用”和“中药多成分单靶点叠加作用”的理论。结论采用高分辨质谱分析技术和网络药理学及其数据库,可以为全面阐释中药(复方)药效物质基础及探讨其分子水平的作用机制提供可行的科学方法。明确了古代经典名方保元汤的入血化学成分,鉴定了其原型成分的代谢产物,并从分子水平阐释了基于这些入血原型成分的作用靶点,并揭示其潜在治疗优势,为扩大该方剂临床应用提供参考,发掘更多的临床治疗潜能。
Objective To elucidate the pharmacodynamic substances and the molecular mechanism of the ancient classical formula Baoyuan Decoction(BYT),in order to explore more potential clinical application advantages of BYT.Methods The UPLC-QTOF-MS/MS technology was was used to perform gradient elution on a Waters ACQUITY UPLC-HSS T3 column(100 mm×2.1 mm,1.7μm)with 0.1%formic acid water(A)-acetonitrile(B)as mobile phase system.High resolution time-of-flight mass spectrometry electrospray ionization source(ESI)and positive and negative ion scanning modes were used to detect and identify the chemical components of BYT,and the prototype chemical components and metabolites in the drug-containing plasma of BYT rats after multiple and multi-day ig administration were analyzed.Furthermore,key targeting effects and the therapeutic potential of the absorbed prototype components of BYT into blood were predicted using network pharmacological methods through TCMSP database and drug information database.Results According to accurate relative molecular mass data and multistage mass spectrometry fragment ions,combined with reference products,databases and literature reports,a total of 133 chemical components in BYT were identified,and it was determined that 35 components could be absorbed into the blood through oral absorption to achieve stable plasma concentration after rats were given BYT by ig.The“component-target”network map of the absorbed prototype components of BYT into blood,core target gene gene ontology(GO)functional enrichment analysis,Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis,and disease prediction analysis indicated that BYT might act on mitogen-activated protein kinase 1(MAPK1),tumor protein p53(TP53),signal transducer and activator of transcription and other core targets through eight key components,including ginsenoside Rb1,paeonol,formononetin,echinatin,daidzein,licochalcone A,6-gingerol and salicylic acid,and exert medicinal effects through the signaling pathways of advanced glycation end products-receptor for advanced glycation end-products(AGE-RAGE),epidermal growth factor receptor(EGFR),janus kinase-signal transducer and activator of transcription(JAK-STAT).It suggests that BYT has many potential therapeutic advantages on such as Alzheimer disease,reperfusion injury,postmenopausal osteoporosis,besides chronic heart failure.There were several derivatives of the same components in the 35 absorbed prototype components of BYT,,and one compound can dock with multiple targets,as well multiple analogues dock with the same target,which supporting the theoretical understanding of“multi-component,multi-target synergy of traditional Chinese medicine(formula)”and“multi-component,single-target superposition of traditional Chinese medicine”.Conclusion Applying the UPLC-QTOF-MS/MS,network pharmacological and its database,which could provide a feasible scientific method for exploring the pharmacodynamic substances of traditional Chinese medicine(formula)and its molecular mechanism of action,as well clarifying the absorbed prototype components of BYT into blood and identifying some metabolites of some prototype components.And the target of action based on these prototype components was explained from the molecular level,and its potential therapeutic advantages were revealed,so as to provide reference for expanding the clinical application of this prescription and explore more clinical therapeutic potential.
作者
李淑慧
王雅
田军
郭雨轩
张淑涵
牟越
翁小刚
孙奕
LI Shu-hui;WANG Ya;TIAN Jun;GUO Yu-xuan;ZHANG Shu-han;MU Yue;WENG Xiao-gang;SUN Yi(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Life Sciences and Engineering,Lanzhou University of Technology,Lanzhou 730050,China;JiangSu Wanbang Biopharmaceuticals,Xuzhou 221001,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第21期6971-6987,共17页
Chinese Traditional and Herbal Drugs
基金
中国中医科学院科技创新工程(CI2021A04514)。
