基于LC-MS代谢组学技术的柴芩宁神颗粒治疗肝郁失眠证的作用机制分析  

Mechanism of Chaiqin Ningshen Granule(柴芩宁神颗粒)in the Treatment of Insomnia with Ganyu(肝郁)Symptom Based on LC-MS Metabolomics

作　　者：赵琳儒 张月 何盼 武英茹 蒋境兴 李杰[3] 张雅筠 刘聪[3] 郝旭亮 Zhao Linru;Zhang Yue;He Pan;Wu Yingru;Jiang Jingxing;Li Jie;Zhang Yajun;Liu Cong;Hao Xuliang(Shanxi University of Chinese Medicine,Jinzhong 030619;?Shanxi University,Taiyuann 030006;Shanxi Academy of Chinese Medicine,Taiyuan 030012;The Hospital of Shanxi University of Chinese Medicine,Taiyuan 030024)

机构地区：[1]山西中医药大学,晋中中030619 [2]山西大学,太原030006 [3]山西省中医药研究院,太原030012 [4]山西中医药大学附属医院,太原030024

出　　处：《中药药理与临床》2022年第6期64-70,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：山西省中医药研究院中药新药研究专项(编号:2016XYZX02);中药新产品关键技术开发实验室(编号:2020SYS09)。

摘　　要：目的:基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q Exactive Orbitrap-MS)代谢组学技术,探讨柴芩宁神颗粒对肝郁失眠大鼠的作用及可能的作用机制。方法:采用注射对氯苯丙氨酸(PCPA)、夹尾和束缚前肢的方法建立符合中医肝郁失眠特征的大鼠模型,观察各组大鼠精神和行为状况,体质量变化,并在试验结束后进行全血粘度、血清5-羟色胺(5-HT)、γ-氨基丁酸(GABA)、谷氨酸(GLU)含量检测;通过代谢组学技术对大鼠血清进行检测,并采用主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)及正交偏最小二乘法判别分析(OPLS-DA)等多元统计分析方法,获取各组大鼠血清中的差异代谢物,通过MetPA数据库寻找相关的代谢途径。结果:与正常对照组比较,模型对照组大鼠的精神萎靡不振,肝郁血瘀现象严重,血清5-HT、GABA含量显著降低,GLU含量显著升高(P<0.01);与模型对照组比较,各给药组大鼠的精神状态和肝郁血瘀现象被改善,艾司唑仑0.5 mg/kg组、柴芩宁神颗粒1.12、2.25 g/kg组血清中5-HT、GABA含量显著升高、GLU含量显著降低(P<0.01),0.56 g/kg组中GABA含量显著升高(P<0.01);通过LC-MS代谢组学技术,从血清中筛选出与失眠相关的46种差异代谢物,其中,艾司唑仑5 mg/kg组可显著回调2种差异代谢物,柴芩宁神颗粒2.25 g/kg组可回调5种差异代谢物,色氨酸代谢、甘油磷脂代谢、组氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢、精氨酸和脯氨酸代谢为药物干预失眠的代谢途径。结论:柴芩宁神颗粒对治疗失眠有一定的疗效,且柴芩宁神颗粒调节的通路较艾司唑仑更多,体现了中药及其复方作用的多途径、多靶点特点,为柴芩宁神颗粒临床应用提供理论依据。Objective:To explore the effect of Chaiqin Ningshen granule(柴芩宁神颗粒)on insomnia rats with Ganyu(肝郁)symptom and its possible mechanism based on ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Q Exactive Orbitrap-MS).Methods:The model of insomnia with Ganyu symptom was induced in rats by injecting p-chlorophenylalanine(PCPA),clamping tail,and binding forelimbs,and the mental and behavioral status and weight changes of rats in each group were observed.The whole blood viscosity,5-hydroxytryptamine(5-HT),γ-aminobutyric acid(GABA),and glutamate(GLU)were determined after the experiment.The serum of rats was determined by metabolomics technology,and multivariate statistical analysis methods such as principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to obtain the differential metabolites in the serum of rats in each group.The relevant metabolic pathways were screened out from MetPA database.Results:As compared with the blank control group,rats in the model control group were depressed with serious Ganyu(肝郁)Xueyu(血瘀)symptom.The levels of 5-HT and GABA decreased significantly,and the level of GLU increased significantly in the model control group.After administration,the mental state,Ganyu symptom,and blood stasis of rats in each administration group were improved as compared with the model control group.The content of 5-HT,GABA,and GLU in serum in the 0.5 mg/kg estazolam group,and the 2.25 g/kg and 1.125 g/kg Chaiqin Ningshen granule groups,and the content of GABA in the 0.5625 g/kg Chaiqin Ningshen granule group were significantly improved.The therapeutic effects of the 0.5 mg/kg estazolam group and the 2.25 g/kg Chaiqin Ningshen granule group were superior.Through LC-MS metabolomics technology,46 differential metabolites related to insomnia were screened out from serum,of which 29 differential metabolites could

关 键 词：柴芩宁神颗粒 肝郁 失眠 代谢组学 潜在生物标志物 代谢通路 

分 类 号：R285.5[医药卫生—中药学]