该实验在已经分离鉴定出牛乳源促睡眠肽的基础上，研究了酸枣仁、百合、睡眠肽3种物质复配制成的复合物对DL-4-氯苯基丙氨酸（PCPA）致失眠大鼠的睡眠干预效果及作用机制。构建PCPA致大鼠失眠模型，通过旷场试验（OFT）探究大鼠的行为学变化，探究复合物对失眠大鼠的促睡眠效果。通过检测大鼠脑部的神经递质及代谢产物研究复合物的作用途径，探究复合物的促睡眠机理。结果显示，复合物组大鼠在中央停留次数、中央区停留时间、运动总路程分别为11.00次、15.01 s和3309.71 cm，与失眠模型组有显著差异（p<0.05），与空白组无显著差异（p>0.05）。复合物组GABA（γ-氨基丁酸）、Glu（谷氨酸）、Gly（甘氨酸）、NE（去甲肾上腺素）、DA（多巴胺）和5-HT（五羟色胺）含量分别为94.96 μg/mL、1508.42 μg/L、74.59 μg/L、37.36 μg/L、1.76 μg/L和1.61 μg/L，复合物组与失眠模型组相比，大鼠下丘脑抑制性神经递质GABA含量显著上升（p<0.05）；兴奋性神经递质NE、DA含量显著下降（p<0.05），表明复合物组可调节大鼠下丘脑中的神经递质。结果说明酸枣仁百合睡眠肽复合物能改善失眠大鼠行为学与睡眠。

The effect and mechanism of action of a mixture of sleep-inducing peptides isolated from cow milk and Semen ziziphi spinosae and Bulbus lilii extracts on para-chlorophenylalanine-induced (PCPA-induced) insomnia in rat models were investigated. The sleep-inducing effect of the mixture was evaluated by studying the behavioral changes of the ratsusingopen-field tests. By measuring the levels of neurotransmitters and metabolites in the rat brains, the reaction pathway and sleep-inducing mechanism of the mixture were analyzed. There were significant differences between rats treated with the mixture and those in which insomnia had been induced (p<0.05) but no significant differences between the treatment and control groups (p>0.05). The number of stays in the central zone, the duration of the stays, and the total distances movedby the rats treated with the mixture were 11.00, 15.01 s, and 3309.71 cm, respectively. The concentrations of γ-aminobutyric acid (GABA), glutamic acid (Glu), glycine (Gly), noradrenaline (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the brains of the mixture-treated group were 94.96 μg/mL, 1508.42 μg/L, 74.59 μg/L, 37.36 μg/L, 1.76 μg/L, and 1.61 μg/L, respectively. In addition, the mixture-treated rats had significantly higher levels of hypothalamic inhibitory neurotransmitters, namely GABA, than the rats with insomnia (p<0.05) but significantly lower levels of excitatory neurotransmitters, namely NE and DA (p<0.05). The results suggest that the mixture of cow-milk-derived sleep-inducing peptides and Semen ziziphi spinosae and Bulbus lilii extracts can regulate the release of hypothalamic neurotransmittersand improve the behavior and sleep of rat models of insomnia.

中国博士后面上项目（2020M672651）；国家自然科学基金项目（31972078）；广东省功能食品活性物重点实验室（2018B030322010）；广东省现代农业产业技术体系创新团队项目（2020KJ117）；广东高校国际科技合作创新平台项目（2013gjhz0003）