宁心安神方调控失眠大鼠Glu/GABA-Gln代谢环路失衡的机制研究

目的研究宁心安神方对失眠的治疗作用及对失眠大鼠谷氨酸(Glu)/γ-氨基丁酸(GABA)-谷氨酰胺(Gln)代谢环路的调控机制。方法采用改良多平台水环境睡眠剥夺法建立失眠模型。将大鼠随机分为正常对照组,模型组,地西泮组,宁心安神方低、中、高剂量组。观察大鼠一般情况,利用戊巴比妥钠睡眠协同实验观察大鼠的睡眠潜伏时间和睡眠持续时间,进行模型评价及宁心安神方治疗失眠的疗效评价。高效液相色谱法检测脑干、下丘脑内神经递质Glu、GABA的含量。利用Western blot法对脑干、下丘脑内谷氨酸脱羧酶(GAD)、星形胶质细胞谷氨酰胺合成酶(GS)的表达水平进行测定。结果宁心安神方中、高剂量可以明显缩短失眠大鼠睡眠潜伏时间、延长睡眠持续时间,提高脑干、下丘脑GABA水平,降低Glu/GABA的比值;宁心安神方各剂量均可明显升高脑干GAD表达和下丘脑中GS的表达。结论宁心安神方的镇静催眠作用可能是通过调控Glu/GABA-Gln代谢环路,影响Glu、GABA的合成和分解,进而改善Glu/GABA的兴奋/抑制代谢失衡而实现的。

Mechanism of Ningxin Anshen Fang in regulation of unbalanced Glu/GABA-Gln metabolic loop in insomnia rats

Objective To study the therapeutic effect of Ningxin Anshen Fang on insomnia, and regulative mechanism to the metabolic loop of glutamic acid (Glu)/γ-aminobutyric acid (GABA)-glutamine (Gln).Methods The model of insomnia was established by using modified multiple water platform sleep deprivation method in rats, and then all rats were randomly divided into normal group, model group, diazepam group, Ningxin Anshen Fang low-dose group (low-dose group), Ningxin Anshen Fang mid-dose group (mid-dose group) and Ningxin Anshen Fang high-dose group (high-dose group).The general conditions were observed, sleep latency and sleep duration were observed by applying pentobarbital sodium (PS) cooperative sleep test, and model and curative effect of Ningxin Anshen Fang on insomnia were reviewed.The content of Glu and GABA in brainstem and hypothalamus was detected by using high performance liquid chromatography (HPLC).The expressions of glutamic acid decarboxylase (GAD) and astrocyte glutamine synthetase (GS) in brainstem and hypothalamus were detected by using Western blot experiment.Results The sleep latency was significantly shortened, sleep duration was prolonged, GABA level in brainstem and hypothalamus was increased and ratio of Glu to GABA (Glu/GABA) was decreased by Ningxin Anshen Fang in insomnia rats.The expression of GAD in brainstem and hypothalamus was significantly promoted in all Ningxin Anshen Fang groups.Conclusion Ningxin Anshen Fang can affect the synthesis and decomposition Glu and GABA and expressions of Glu and GABA, and normalize the excitotary/inhibitory metabolic imbalance of Glu/GABA through regulating Glu/GABA-Gln metabolic loop, which may be related to that Ningxin Anshen Fang has the curative effects of calming heart and tranquillizing mind, and sedation and hypnosis.