基于^(1)H NMR代谢组学与网络药理学的复方宁神精油抗失眠组分物质基础及作用机制研究  

Study on the material basis and mechanism of anti-insomnia mechanism of Ning Shen Essential Oil based on ^(1)H NMR metabolomics and network pharmacology

作　　者：柴青 张宏斌 吴立东 王婧毅 李海超 刘玉红[1,5,6] 刘红燕 蒋海强[2,5,6] 田振华 CHAI Qing;ZHANG Hong-bin;WU Li-dong;WANG Jing-yi;LI Hai-chao;LIU Yu-hong;LIU Hong-yan;JIANG Hai-qiang;TIAN Zhen-hua(School of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Institute of Traditional Chinese Medicine Innovation,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Shandong Xinyue Health Science and Technology Co.,Ltd,Binzhou 256600,China;Experimental Center,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Shandong Provincial Key Laboratory of Basic Research of Traditional Chinese Medicine,Jinan 250355,China;Key Laboratory of Classical Theory of Chinese Medicine,Ministry of Education,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]山东中医药大学中医药创新研究院,山东济南250355 [3]山东欣悦健康科技有限公司,山东滨州256600 [4]山东中医药大学实验中心,山东济南250355 [5]山东省中医药基础研究重点实验室,山东济南250355 [6]山东中医药大学,教育部中医药经典理论重点实验室,山东济南250355

出　　处：《药学学报》2024年第8期2313-2325,共13页Acta Pharmaceutica Sinica

基　　金：山东省青创人才引育计划。

摘　　要：本文应用GC-MS、网络药理学与^(1)H NMR代谢组学技术相结合,研究复方宁神精油抗失眠的物质基础与作用机制。通过GC-MS分析复方宁神精油的主要挥发性成分,利用中药系统药理学数据库与分析平台(TCMSP)及GeneCards、OMIM、Drugbank等数据库预测失眠相关靶点。腹腔注射对氯苯丙氨酸(PCPA)复制大鼠失眠模型。动物实验经山东中医药大学动物伦理委员会批准(伦理号:SDUTCM20221025010)。以行为学相关指标评价复方宁神精油对大鼠焦虑行为的调节作用;采用酶联免疫法检测大鼠血清中促肾上腺激素释放激素(CRH)、促肾上腺激素(ACTH)、褪黑素(MT)含量;取大鼠血清、海马进行^(1)H NMR代谢组学检测大鼠血清海马中内源性代谢物变化,指认差异代谢物并构建代谢通路。结果显示,模型组大鼠旷场实验运动距离、高架十字迷宫实验进入开放臂次数及时间均显著降低(P<0.05,P<0.01);复方宁神精油干预后,大鼠行为学指标均不同程度改善。模型组大鼠血清CRH、ACTH含量均显著升高(P<0.05,P<0.01),MT含量显著降低(P<0.01);干预后,血清CRH、ACTH含量均不同程度降低,且能回调MT含量。^(1)H NMR代谢组学共筛选10种与失眠相关的潜在生物标志物,涉及到6条潜在的代谢通路。通过GC-MS共检测出复方宁神精油35个成分,复方宁神精油主要成分靶点与失眠疾病靶点取交集,共筛选出172个交集基因,核心靶点26个。研究表明复方宁神精油干预PCPA所致失眠大鼠的作用机制可能与调节下丘脑-垂体-肾上腺轴轴相关激素,调控机体氨基酸代谢、脂质代谢和胆碱代谢有关。This paper applied gas chromatography-mass spectrometry(GC-MS),network pharmacology and nuclear magnetic resonance hydrogen spectroscopy(^(1)H NMR)metabolomics techniques to study the material basis and mechanism of action of Ning Shen Essential Oil in anti-insomnia.The main volatile components of Ning Shen Essential Oil were analyzed by gas chromatography-mass spectrometry(GC-MS),and the insomnia-related targets were predicted using the Traditional Chinese Medicine Systematic Pharmacology Database and Analytical Platform(TCMSP)and the databases of GeneCards,OMIM and Drugbank.The insomnia model of rats was replicated by intraperitoneal injection of 4-chloro-DL-phenylalanine(PCPA).Animal experiments were approved by the Animal Ethics Committee of Shandong University of Traditional Chinese Medicine(Ethics No.:SDUTCM20221025010).The modulating effect of Compound Ning Shen Essential Oil on anxiety behavior of rats was evaluated by behavioral related indexes.The serum levels of corticotropin-releasing hormone(CRH),adrenotropic corticotropic hormone(ACTH)and melatonin(MT)were measured by enzyme-linked immunoassay(ELISA).Rat serum and hippocampus were taken for nuclear magnetic resonance(^(1)H NMR)metabolomics to detect the changes of endogenous metabolites in rat serum hippocampus,to designate the differential metabolites and to construct metabolic pathways.The results showed that the exercise distance in the open field experiment and the number of times and time to enter the open arm in the elevated cross maze experiment of the rats in the model group were significantly reduced(P<0.05,P<0.01).The behavioral indexes of rats improved to different degrees after the administration of Ning Shen Essential Oil.The serum CRH and ACTH levels of rats in the model group increased significantly(P<0.05,P<0.01),and the MT level decreased significantly(P<0.01);After the intervention,serum CRH and ACTH levels were reduced to different degrees,and MT levels could be regressed.^(1)H NMR metabolomics screened 10 potential biomarkers rel

关 键 词：复方宁神精油 失眠 旷场实验 高架十字迷宫 核磁共振 代谢组学 网络药理学 

分 类 号：R917[医药卫生—药物分析学]