酸枣仁-茯苓-党参水提物对小鼠睡眠的改善作用及机制研究

目的:探究酸枣仁-茯苓-党参水提物（质量比1:1:1）对小鼠睡眠的改善作用，并探讨其潜在作用机制。方法:120只雄性昆明小鼠中选取60只随机分为6组，进行自主活动实验及戊巴比妥钠诱导睡眠实验，分为正常组、溶剂对照组、酸枣仁-茯苓-党参低、中、高剂量组（14、28、56 g/kg）、阳性药组（地西泮，2.5 mg/kg）；另外60只小鼠进行对氯苯丙氨酸诱导失眠实验，分为正常组、模型组、酸枣仁-茯苓-党参低、中、高剂量组、阳性药组，每组10只，灌胃剂量为10 mL/（kg·d），不间断给药30 d。通过记录小鼠自主活动，分析水平得分与活动距离，评价酸枣仁-茯苓-党参水提物对正常小鼠及对对氯苯丙氨酸建立的失眠小鼠的的影响；通过戊巴比妥钠诱导睡眠实验，记录小鼠睡眠潜伏期和睡眠时间，评价酸枣仁-茯苓-党参水提物的促进睡眠及延长睡眠时间的作用。ELISA检测失眠实验中各组小鼠下丘脑、皮层中5-羟色胺（5-HT）及色氨酸羟化酶（TPH）的含量；小鼠海马、皮层中谷氨酸（Glu）及γ-氨基丁酸（GABA）的含量，探讨酸枣仁-茯苓-党参水提物相关作用机制。结果:酸枣仁-茯苓-党参水提物能够显著减少小鼠睡眠潜伏期，延长睡眠时间，降低失眠小鼠自主活动能力，减少正常小鼠水平得分与活动距离（P < 0.05，P<0.01）；高剂量酸枣仁-茯苓-党参水提物能够增强下丘脑5-HT合成关键酶TPH的活性，提高5-HT含量（P<0.05）；能够调节海马Glu与GABA含量，维持Glu/GABA平衡（P<0.05）。结论:酸枣仁-茯苓-党参水提物对睡眠起到一定的改善作用，作用机制与调节小鼠脑内血清素系统和Glu/GABA稳态有关。

Effects of Water Extract of Ziziphi Spinosae Semen, Poria Cocos and Codonopsis Radixon on Sleep Function in Mice and Its Underlying Mechanisms

Objective: To explore the effect of water extract of Ziziphi Spinosae Semen, Poria Cocos and Codonopsis Radixon (1:1:1) on sleep improvement in mice and its potential mechanism. Methods: 60 male KM mice were randomly divided into 6 groups: normal group, solvent control group, Ziziphi Spinosae Semen-Poria cocos-Codonopsis Radixon low, medium, high dose group (14, 28, 56 g/kg), positive drug group (diazepam, 2.5 mg/kg), autonomous activity experiments and pentobarbital induced sleep experiments were performed; another 60 KM mice were tested for insomnia induced by p-chlorphenylalanine, It was divided into normal group, model group, Ziziphi Spinosae Semen-Poria cocos-Codonopsis Radixon low, medium, high dose group, positive drug group, 10 in each group, gavage dose of 10 mL/(kg·d), continuous administration for 30 d. The sleep latency and time were recorded by sodium pentobarbital, and the effect of water extract on promoting sleep and prolonging sleep time was evaluated. By recording the autonomous activity of mice, analyzing the score of level and the distance of activity, we evaluated the effect of water extract on the improvement of PCPA induced insomnia mice and the effect on normal mice. Contents of 5-HT and TPH in hypothalamus and cortex, Glu and GABA in hippocampus and cortex of mice were detected. Results: The extract could significantly reduce the sleep latency, prolong the sleep time, reduce the autonomous activity ability of insomnia mice, and reduce the level score and activity distance of normal mice (P < 0.05, P< 0.01); enhance the activity of 5-HT synthesis key enzyme TPH, increase the content of 5-HT in hypothalamus (P< 0.05); regulate the content of hippocampal Glu and GABA and maintain the Glu/GABA balance (P< 0.05). Conclusion: The water extract of Ziziphi Spinosae Semen, Poria Cocos and Codonopsis Radixon can improve sleep, and the mechanism is related to regulating the serotonin system and Glu/GABA homeostasis in the mouse brain.