眠得乐方调控神经递质改善失眠大鼠睡眠行为的研究

Study on the Regulation of Neurotransmitters to Improve Sleep Behavior in Insomnia Rats

目的观察“眠得乐方”对失眠大鼠的行为学、脑组织及中枢神经递质的影响。方法将SD雄性大鼠随机分为对照组、模型组、眠得乐方组、艾司唑仑组,每组10只,采用腹腔注射氯苯丙氨酸(Pchlorophenylalanine,PCPA)的方法构建失眠模型。造模成功后,对照组与模型组生理盐水灌胃,眠得乐方组与艾司唑仑组分别给予药物灌胃。通过戊巴比妥钠翻正实验和旷场实验评价各组大鼠失眠样行为,苏木精-伊红(Hematoxylin and eosin,HE)染色观察大鼠大脑皮质、海马、下丘脑组织的病理改变,酶联免疫吸附测定法(Enzyme-linked mmunosorbent assay,ELISA)测定大鼠大脑皮质、下丘脑、海马区的γ-氨基丁酸(Gamma-aminobutyric acid,GABA)、5-羟色胺(5-hydroxytryptamine,5-HT)的表达水平。结果模型组大鼠的睡眠潜伏期显著拉长(P<0.01),而睡眠时间较少(P<0.01),精神状态、皮毛色泽等较差,体质量明显减轻(P<0.01),与模型组比较,眠得乐方组和艾司唑仑组大鼠的睡眠潜伏期显著缩短(P<0.01),睡眠持续时间显著拉长(P<0.01),体质量显著升高(P<0.05,P<0.01);旷场实验中模型组大鼠总活动距离增加,平均速度和中心区域停留时间减少(P<0.05),眠得乐方组和艾司唑仑组大鼠总活动距离显著减少(P<0.05),平均速度增加和中心区域停留时间延长(P<0.05)。HE结果显示模型组大鼠大脑皮质、海马、下丘脑神经元数量、形态结构、排列等均有不同程度受损,眠得乐方组及艾司唑仑组显著改善。ELISA结果显示模型组大鼠大脑皮质和下丘脑的5-HT和GABA表达量显著降低(P<0.01,P<0.05),海马组织的GABA表达量亦显著降低(P<0.01)。眠得乐方组大脑皮质GABA和下丘脑GABA、5-HT的蛋白表达量均显著升高(P<0.01)。结论眠得乐方能够改善失眠大鼠的精神状态、恢复正常体质量、提高睡眠效率、减轻焦虑和紧张度。其机制可能与升高5-HT和GABA的含量,抑制下丘脑和脑干促觉醒核团的扩散及传导有关。

Objective To observe the effects of"Sleep Recipe"on the behavior,brain tissue and central neurotransmitters of insomnia rats.Methods The male rats with SD were randomly divided into control group,model group,sleep formula group,and eszolam group,with 10 rats in each group,and the insomnia model was constructed by intraperitoneal injection of P-chlorphenylalanine(PCPA).After successful modeling,the control group and the model group were given saline gavage,and the medylom group and eszolam group were given drug gavage.The insomnia-like behavior of rats in each group was evaluated by pentobarbital sodium correction experiment and open field experiment,Hematoxlin and eosin(HE)staining observed the pathological changes of rat cerebral cortex,hippocampus,and hypothalamic tissue,and Enzyme-linked mmunosorbent assay(ELISA)was used to determine the expression levels of Gamma-aminobutyric acid(GABA),5-hydroxytryptamine(5-HT).Results The sleep latency of rats in the model group was significantly elongated(P<0.01),while the sleep time was less(P<0.01),and the mental state and fur color were poor,significantly decreased in body weight(P<0.01).Compared with the model group,the sleep latency was significantly shortened(P<0.01),the sleep duration was significantly prolonged(P<0.01),and the body mass was significantly increased(P<0.05,P<0.01);In the open field experiment,the total activity distance of rats in the model group increased,the average speed and central region residence time decreased(P<0.05),the total activity distance of rats in the Sleep Formula group and Eszolam group decreased significantly(P<0.05),the average speed increased and the central region residence time increased(P<0.05).HE results showed that the number of neurons,morphological structure and arrangement of neurons,such as cerebral cortex,hippocampus and hypothalamus in the model group,were damaged to varying degrees,and the sleep formula group and estazolam group were significantly improved.ELISA results showed that the expression of 5-HT and GABA in the cerebral cortex and hypothalamus of rats in the model group was significantly reduced(P<0.01,P<0.05),and the expression of GABA in hippocampal tissues was also significantly reduced(P<0.01).The protein expression levels of cerebral cortical GABA,hypothalamic GABA and 5-HT in the Sleep Formula group were significantly increased(P<0.01).Conclusion Sleep Formula can improve the mental state,restore normal body weight,improve sleep efficiency,and reduce anxiety and tension in insomnia rats.The mechanism may be related to increasing the content of 5-HT and GABA,and inhibiting the spread and conduction of hypothalamic and brainstem pro-awakening nuclei.