目的 利用代谢组学技术探究生酸枣仁（Ziziphi Spinosae Semen，ZSS）和炒酸枣仁（fried Ziziphi Spinosae Semen，FZSS）对失眠大鼠内源性代谢物的影响，并进一步分析其对学习记忆功能的改善效果及作用机制。方法 通过对氯苯丙氨酸（p-chlorophenylalanine，PCPA）诱导建立大鼠失眠模型，给予ZSS和FZSS干预7 d。通过旷场实验和Morris水迷宫实验评估大鼠的学习行为，利用ELISA测定海马组织中5-羟色胺（5-hydroxytryptamine，5-HT）、γ-氨基丁酸（γ-aminobutyric acid，GABA）、一氧化氮（nitric oxide，NO）和血清中白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、IL-1β、褪黑素（melatonin，MT）、谷氨酸（glutamic acid，Glu）水平。采用超高效液相色谱-串联质谱（UPLC-Orbitrap-MS/MS）技术对大鼠海马和血清样本的代谢轮廓进行详细分析，并结合多元统计方法筛选出内源性差异性代谢物。通过MetaboAnalyst 5.0数据库进行代谢通路和网络的分析，多角度探讨ZSS和FZSS对学习记忆功能改善的代谢特征及其作用机制。结果 与模型组比较，ZSS组和FZSS组大鼠的学习记忆表现显著提升，Morris水迷宫实验中目标象限停留时间、穿越次数及运动路程均显著增加（P＜0.05、0.01）；海马组织中5-HT、GABA和NO水平显著升高（P＜0.05、0.01、0.001），其中FZSS对GABA的回调效果更显著；血清中IL-6、TNF-α、Glu水平显著降低（P＜0.05、0.01、0.001），IL-1β、MT水平显著升高（P＜0.05、0.01），海马神经元空化等病理损伤得到显著改善。ZSS和FZSS显著回调海马相关代谢物34个以及血清中相关代谢物22个，主要调控苯丙氨酸、酪氨酸和色氨酸3个代谢通路，且FZSS特异性调控鞘脂代谢通路，ZSS特异性调控花生四烯酸代谢通路。结论 ZSS和FZSS通过调节海马和血清中的内源性代谢物（尤其是苯丙氨酸、酪氨酸、色氨酸通路），显著改善PCPA诱导失眠导致的学习记忆损伤。炒制使FZSS中木兰花碱、斯皮诺素、酸枣仁皂苷A等有效成分含量增加，增强其对鞘脂代谢的调控及GABA水平的回调能力，因此在改善认知损伤方面优于ZSS。为酸枣仁及其炮制品治疗失眠相关认知障碍提供了代谢水平的科学依据，为临床治疗提供一种新的治疗策略。

Objective To explore the effects of Suanzaoren (Ziziphi Spinosae Semen, ZSS) and fried Ziziphi Spinosae Semen (FZSS) on endogenous metabolites in insomnia rats using metabolomics technology. In addition, the potential improvement effect and its mechanism on learning and memory function were further analyzed. Methods The rat insomnia model was induced by p-chlorophenylalanine (PCPA), ZSS and FZSS were given for 7 d. The learning behavior of rats was evaluated by open field test and Morris water maze test; The levels of 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), nitric oxide (NO) in hippocampus and interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), IL-1β, melatonin (MT), and glutamic acid (Glu) in serum were measured by ELISA. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-Orbitrap-MS/MS) was used to analyze the metabolic profiles of rat hippocampus and serum samples in detail, and multivariate statistical methods were used to screen out differential metabolites. The metabolic pathway and network were analyzed by MetaboAnalyst 5.0 database to explore the metabolic characteristics and mechanism of ZSS and FZSS in improving learning and memory function from multiple perspectives. Results Compared with model group, rats in ZSS and FZSS groups showed significantly improved learning and memory performance. In the Morris water maze test, time spent in the target quadrant, number of platform crossings and total path length were significantly increased (P < 0.05, 0.01), levels of 5-HT, GABA and NO in hippocampal tissue were significantly increased (P < 0.05, 0.01, 0.001), with FZSS demonstrating a more significant restorative effect on GABA level; The levels of IL-6, TNF-α and Glu in serum were significantly decreased (P < 0.05, 0.01, 0.001), levels of IL-1β and MT were significantly increased (P < 0.05, 0.01), pathological damage such as neuronal vacuolation in hippocampus was ameliorated. ZSS and FZSS significantly restored 34 relevant metabolites in hippocampus and 22 relevant metabolites in serum, primarily regulated three metabolic pathways, which are phenylalanine, tyrosine, and tryptophan biosynthesis and metabolism. Notably, FZSS specifically regulated the sphingolipid metabolism pathway, while ZSS specifically regulated the arachidonic acid metabolism pathway. Conclusion ZSS and FZSS significantly ameliorate learning and memory impairments induced by PCPA insomnia through regulating endogenous metabolites in hippocampus and serum, particularly within the phenylalanine, tyrosine, and tryptophan pathways. The stir-frying process increased the contents of active components such as magnoflorine, spinosin and jujuboside A in FZSS, which enhanced its regulatory effect on sphingolipid metabolism and its restorative capacity on GABA level. Consequently, FZSS demonstrates superior efficacy over the raw ZSS in improving cognitive impairment. This study provides a scientific basis at the metabolic level for using ZSS and its processed product (FZSS) to treat insomnia-related cognitive disorders and offers a novel therapeutic strategy for clinical application.

R285.5

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