基于血清代谢组学探讨复方枣仁颗粒改善失眠的机制  

Investigation on the mechanism of Compound zaoren granules in improving insomnia based on serum metabonomics

作　　者：王泽琨 刘沈林 于小聪 李丹婷 张凌峰 赵毅萌 陈辰[3] 陈亚军[3] 束雅春 WANG Zekun;LIU Shenlin;YU Xiaocong;LI Danting;ZHANG Lingfeng;ZHAO Yimeng;CHEN Chen;CHEN Yajun;SHU Yachun(Dept.of Pharmacy,the Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;School of Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 211198,China;Dept.of Clinical Laboratory,Nanjing Maternal and Child Health Care Hospital,Nanjing 210001,China)

机构地区：[1]南京中医药大学附属医院药学部,南京210029 [2]中国药科大学中药学院,南京211198 [3]南京市妇幼保健院检验科,南京210001

出　　处：《中国药房》2023年第9期1093-1098,共6页China Pharmacy

基　　金：南京市科技计划项目(No.202002054);江苏省研究生科研与实践创新计划项目(No.SJCX21_0669)。

摘　　要：目的研究复方枣仁颗粒改善失眠的机制。方法将49只小鼠随机分为空白组、模型组、阳性对照组1(艾司唑仑片0.5 mg/kg)、阳性对照组2(舒眠胶囊0.6 g/kg)和复方枣仁颗粒低、中、高剂量组(2.5、5、10 g/kg),每组7只。通过慢性不可预见性温和应激刺激联合对氯苯丙氨酸建立小鼠失眠模型,通过小鼠旷场实验、戊巴比妥钠协同催眠实验考察小鼠行为学变化,通过苏木精-伊红染色观察小鼠下丘脑组织病理形态学变化。结合超高效液相色谱-飞行时间质谱技术对小鼠血清进行代谢组学分析及多元统计分析筛选差异代谢物,并基于MetaboAnalyst 5.0数据库进行代谢通路分析。结果与空白组比较,模型组小鼠总行进距离、中央区域进入次数、中央区域移动距离显著减少(P<0.05),总静息时间比例显著升高(P<0.05),睡眠时长显著缩短(P<0.05),下丘脑神经细胞受损且空泡化严重。与模型组比较,复方枣仁颗粒低、中剂量组小鼠总行进距离显著增加(P<0.05)、总静息时间比例显著降低(P<0.05),复方枣仁颗粒高剂量组小鼠睡眠时长显著延长(P<0.05);各给药组小鼠下丘脑组织神经细胞均不同程度恢复,其中复方枣仁颗粒高剂量组小鼠下丘脑组织细胞与空白组较为接近。通过代谢组学分析共得到18个差异代谢物(如苯基丙氨酸、牛磺酸、正缬氨酸、甲硫氨酸等)及4条重要的氨基酸代谢通路(L-苯丙氨酸、酪氨酸和色氨酸生物合成,牛磺酸和次牛磺酸代谢,L-苯丙氨酸代谢,半胱氨酸和甲硫氨酸代谢)。结论复方枣仁颗粒可通过回调苯基丙氨酸、牛磺酸等差异代谢物及调节4种氨基酸代谢通路,使紊乱的体内代谢恢复正常,从而发挥改善失眠的作用。OBJECTIVE To study the mechanism of Compound zaoren granule in improving insomnia.METHODS Forty-nine mice were divided into blank group,model group,positive control group 1(Estazolam tablets 0.5 mg/kg),control group 2(Shumian capsule 0.6 g/kg),Compound zaoren granule low-dose,medium-dose and high-dose groups(2.5,5,10 g/kg),with 7 mice in each group.The insomnia model was established by chronic unpredictable mild stress combined with 4-chloro-DLphenylacetic acid.The behavioral changes of mice were investigated through open field test and pentobarbital sodium synergistic hypnosis experiment,as well as the pathomorphology of mice hypothalamus tissue was observed by HE staining.The metabonomics analysis and multivariate statistical analysis of serum in mice were performed by UHPLC-Q-TOF-MS/MS,and the differential metabolites were screened out;the metabolic pathway analysis was conducted based on MetaboAnalyst 5.0 database.RESULTS Compared with blank group,the total travelling distance,the number of entering the central region and the moving distance in the central region of the model group were significantly reduced(P<0.05),the proportion of total rest time was significantly increased(P<0.05),the sleep duration of mice was significantly shortened(P<0.05),and hypothalamic nerve cells damaged and severely vacuolated.Compared with model group,the total travelling distance of Compound zaoren granule low-dose and medium-dose groups were increased significantly and the proportions of total rest time of those groups were decreased significantly(P<0.05),and the sleep duration of mice in Compound zaoren granule high-dose group was prolonged significantly(P<0.05);the hypothalamic nerve cells of mice in each administration group recovered to varying degrees,and the hypothalamus histiocytes of mice in the Compound zaoren granules high-dose group were closer to those in the blank group.A total of 18 differential metabolites(such as phenylalanine,taurine,norvaline,methionine)and 4 important amino acid metabolic pathways(L-phenylal

关 键 词：复方枣仁颗粒 失眠 血清代谢组学 代谢通路 超高效液相色谱-飞行时间质谱 

分 类 号：R285.5[医药卫生—中药学]