中国现代中药2025，Vol.27Issue(4)：695-709,中插6-中插7,17.DOI:10.13313/j.issn.1673-4890.20240819002
基于下丘脑代谢组学的益肾强身丸、防衰益寿丸改善睡眠作用及机制研究
Effects and Mechanisms of Yishen Qiangshen Pills and Fangshuai Yishou Pills on Improving Sleep Based on Hypothalamic Metabolomics
摘要
Abstract
Objective:This study was conducted to investigate the effects and mechanisms of Yishen Qiangshen Pills and Fangshuai Yishou Pills on improving the sleep of mice based on metabolomics technology.Methods:A total of 778-month-old C57BL/6J mice were randomly divided into the control group,model group,Yishen Qiangshen Pills 0.25 g·kg-1 group(YSL),Fangshuai Yishou Pills 0.25 g·kg-1 group(FSL),Yishen Qiangshen Pills+Fangshuai Yishou Pills 0.25 g·kg-1 group(YSFSL),Yishen Qiangshen Pills+Fangshuai Yishou Pills 0.50 g·kg-1 group(YSFSH),and estazolam 0.16 mg·kg-1 group based on their body weight.Middle-aged and elderly mice model with insomnia was prepared through unpredictable stress,and each administration group was administrated at the same time for 8 consecutive weeks.Then,the effect of drugs on sleep in mice was observed under the synergistic effect of sodium pentobarbital with subthreshold dose and threshold dose;elevated plus maze test was performed to evaluate the anxiolytic effect;ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was used to detect neurotransmitter levels in brain tissue;the non-targeted metabolomics method of ultra-high performance liquid chromatography quadrupole electrostatic orbitrap high resolution mass(UHPLC-Q-Exactive Orbitrap/MS)was applied to detect and analyze the hypothalamus;Western blot was further adopted to detect the expression levels of key metabolic enzymes glutamate decarboxylase 1(GAD1)and glutamate decarboxylase 2(GAD2)in the taurine and hypotaurine metabolic pathways.Results:Compared with the model group,mice in the YSL,FSL,YSFSL,and YSFSH groups had shorter latency to sleep and longer sleep duration under the synergistic effect of the threshold dose of sodium pentobarbital(P<0.05,P<0.01);mice in the YSFSL and YSFSH groups had a significantly increased number of times in entering the open arm(P<0.01);the level of γ-aminobutyric acid(GABA)in the brain tissues of mice in the YSL group,the FSL group,the YSFSL group,and the YSFSH group was significantly increased(P<0.001),and glutamate(Glu)level and Glu/GABA ratio were significantly decreased(P<0.05,P<0.01).Metabolomic results showed that the hypothalamic metabolic profile of insomnia mice could be significantly improved after administration,and 12 differential metabolites related to sleep were screened out.Among them,the YSL group could recover 6 differential metabolites,mainly involving metabolic pathways such as taurine and hypotaurine metabolism,purine metabolism,alanine,aspartate,and glutamate metabolism;the FSL group could recover 6 differential metabolites,mainly involving metabolic pathways such as purine metabolism,taurine,and hypotaurine metabolism;the YSFS group could recover 8 differential metabolites,mainly involving metabolic pathways such as taurine and hypotaurine metabolism,alanine,aspartate and glutamate metabolism,citric acid cycle(TCA cycle)and purine metabolism.More importantly,GAD1 and GAD2 were key metabolic enzymes in the taurine and hypotaurine metabolic pathways.Western blot results showed that the expression levels of GAD1 and GAD2 could be significantly reduced after administration.Conclusion:Yishen Qiangshen Pills and Fangshuai Yishou Pills can improve sleep,and the mechanism is related to the regulation of neurotransmitter levels in brain tissue and endogenous metabolites in the hypothalamus.These pills perform a sedative and hypnotic effect by regulating amino acid metabolism,purine metabolism,and energy metabolism in the hypothalamus.关键词
益肾强身丸/防衰益寿丸/镇静/催眠/代谢组学Key words
Yishen Qiangshen pills/Fangshuai Yishou Pills/sedation/hypnosis/metabolomics分类
中医学引用本文复制引用
耿杨,董世芬,冯倩倩,张磊,赵子阳,史浩男,贺润铖,贾占红,张硕峰,孙建宁..基于下丘脑代谢组学的益肾强身丸、防衰益寿丸改善睡眠作用及机制研究[J].中国现代中药,2025,27(4):695-709,中插6-中插7,17.基金项目
国家自然科学基金项目(81503287) （81503287）
