整合代谢组学和肠道菌群探讨生、炒酸枣仁改善睡眠的作用机制特点  

The mechanism of Ziziphi Spinosae Semen and fried Ziziphi Spinosae Semen to improve insomnia based on integrating metabolomics and intestinal flora

作　　者：王丛辉 韩瑞 李泽 衡依然 陈尧楠 裴香萍 闫艳[2] 杜晨晖 WANG Cong-hui;HAN Rui;LI Ze;HENG Yi-ran;CHEN Yao-nan;PEI Xiang-ping;YAN Yan;DU Chen-hui(Institute of Pharmaceutical&Food Engineering,Shanxi University of Chinese Medicine,Taiyuan 030619,China;Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan 030006,China)

机构地区：[1]山西中医药大学中药与食品工程学院,山西太原030619 [2]山西大学中医药现代研究中心,山西太原030006

出　　处：《药学学报》2023年第7期1940-1951,共12页Acta Pharmaceutica Sinica

基　　金：山西省自然科学基金面上项目(20210302123237,20210302123470);山西省科技厅中央引导地方科技发展资金项目(YDZJSX2021C025);山西省教育厅科技创新项目(2020L0422);山西省中医药管理局科研课题(2019ZYYC018);山西中医药大学青年科学家培育项目(2021PY-QN-07);山西中医药大学科技创新团队(2022TD2009).

摘　　要：本研究整合代谢组学技术和肠道菌群分析,探讨生、炒酸枣仁(Ziziphi Spinosae Seme,ZSS;Fried Ziziphi Spinosae Semen,FZSS)对失眠小鼠内源性代谢物及肠道菌群结构的影响。动物实验获得山西中医药大学伦理委员会的批准(批准号:2021DW172)。通过腹腔注射对氯苯丙氨酸(PCPA)复制小鼠失眠模型,分别给予ZSS、FZSS治疗5天,开展戊巴比妥钠诱导睡眠实验,考察ZSS、FZSS协同助眠作用。采用UPLC-Q-Orbitrap MS对失眠小鼠血清进行代谢轮廓分析,结合多元统计分析,筛选差异代谢物,并借助MetaboAnalyst 5.0构建相关代谢通路与网络。利用16S rRNA分析肠道菌群结构及筛选差异性菌群。结果显示,戊巴比妥钠诱导睡眠实验中,与模型组相比,ZSS、FZSS均可降低睡眠潜伏期并延长睡眠时间;血清代谢组学结果显示,共鉴定36种与失眠相关的潜在生物标志物。与模型组相比,ZSS和FZSS组分别鉴定25和27种差异代谢物,其中ZSS、FZSS均可显著回调共有的20种差异代谢物。共涉及10条与氨基酸代谢、能量代谢和脂质代谢调控机制密切相关通路。苯丙氨酸、酪氨酸和色氨酸生物合成,半胱氨酸和甲硫氨酸等5条代谢通路是ZSS组、FZSS组共有调控途径;而花生四烯酸代谢是ZSS改善睡眠的独有代谢途径;谷氨酸和谷氨酰胺等3条代谢通路是FZSS改善睡眠独有的调控途径。PCoA分析表明ZSS组、FZSS组菌群结构均有向空白组恢复的趋势;ZSS、FZSS均可促进普沃菌属丰度下降,而上调乳酸菌属丰度。LEfSe和相关性分析表明血清差异代谢物犬尿喹啉酸、L-谷氨酸、D-脯氨酸与ZSS组差异菌γ-变形菌纲呈显著正相关;5-羟色胺、乙酰胆碱、L-甲硫氨酸与FZSS组差异菌乳酸菌属呈显著正相关。综上,ZSS与FZSS均可影响氨基酸、能量及脂质等代谢通路并调控肠道菌群结构,从而发挥治疗失眠作用,但在具体的代谢通路及肠道菌群各有特点。本研究结果为阐�This study aimed to explore the effects of Ziziphi Spinosae Seme(ZSS)and Fried Ziziphi Spinosae Semen(FZSS)on metabolites and intestinal flora structure in insomnia mice by combining metabolomics analysis and 16S rRNA gene sequencing.Animal experimentation was approved according to the Committee on the Ethics of Animal Experiments of Shanxi University of Chinese Medicine(approval number:2021DW172).The prediction targets were validated in para-chlorophenylalanine(PCPA)induced insomnia mice with administration of ZSS and FZSS for 5 days,respectively.Then pentobarbital sodium induced sleeping test were performed to evaluate the synergistic sleep-aiding effect of ZSS and FZSS.The metabolic profile of serum from insomnia mice was analyzed by UPLC-Q-Orbitrap MS.Different metabolites were screened combined with multivariate statistical analysis.The relevant metabolic pathways and networks were constructed by MetaboAnalyst 5.0.Intestinal flora changes were detected by 16S rRNA sequencing technology.Animal study indicated that,compared to PCPA-induced insomnia model,ZSS and FZSS shortened the sleeping latency and increased the sleeping duration.The serum metabolomics results showed that,there are 36 potential biomarkers associated with insomnia were identified.Compared with the model group,25 and 27 differential metabolites were identified in the ZSS and FZSS groups,respectively.Both ZSS and FZSS groups could significantly adjust to the common 20 differential metabolites.A total of 10 pathways are closely related to insomnia,which including amino acid metabolism,energy metabolism and lipid metabolism.5 metabolic pathways are shared in ZSS and FZSS,including phenylalanine,tyrosine and tryptophan biosynthesis,cysteine and methionine metabolism and so on.Arachidonic acid metabolism is the unique metabolic pathway for ZSS to improve sleep;3 metabolic pathways including glutamate and glutamine are the unique regulatory pathway for FZSS to improve sleep.PCoA analysis showed that the structure of ZSS and FZSS recovered to blank

关 键 词：酸枣仁 炒酸枣仁 代谢组学 肠道菌群 改善睡眠机制 

分 类 号：R917[医药卫生—药物分析学]