基于GABA信号通路探究柏子养心汤对失眠大鼠睡眠时相的影响  

Effects of Baizi Yangxin Decoction on sleep phase in insomnia rats based on GABA signaling pathway

作　　者：谭高峰[1] 乔明亮[1] 郭健[1] 郑伟锋[2] 赵彦青[1] 张希倩 Tan Gaofeng;Qiao Mingliang;Guo Jian;Zheng Weifeng;Zhao Yanqing;Zhang Xiqian(Department of Geriatrics,Encephalopathy District,Zhengzhou 450053,China;Department of Geriatrics,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450053,China)

机构地区：[1]河南省中医院老年病科,河南郑州450053 [2]河南省中医院脑病二区,河南郑州450053

出　　处：《实用药物与临床》2023年第4期293-299,共7页Practical Pharmacy and Clinical Remedies

基　　金：河南省中医药科学研究专项课题(2019ZY2062)。

摘　　要：目的基于γ-氨基丁酸(GABA)信号通路探究柏子养心汤对氯苯丙氨酸(PCPA)致失眠大鼠睡眠时相的影响。方法将SD大鼠分为对照组、模型组、柏子养心汤低剂量、中剂量、高剂量组(5.625 g/kg、11.25 g/kg、22.5 g/kg),每组24只。通过腹腔注射PCPA建立失眠大鼠模型。给予相应剂量的药物连续灌胃7 d后,观察大鼠一般活动状态及体重变化,通过动物睡眠生物解析系统记录脑电图(EEG)和肌电图(EMG)信号,分析大鼠夜晚和白天觉醒(Wake)总量和非快速眼动(NREM)睡眠、快速眼动(REM)睡眠总量;酶联免疫吸附法(ELISA)检测大鼠下丘脑5-羟色胺(5-HT)、谷氨酸(Glu)与GABA含量;免疫组织化学法(IHC)检测大鼠下丘脑谷氨酸脱羧酶67(GAD67)表达;Western blot和RT-qPCR检测大鼠下丘脑组织中γ-氨基丁酸A型(GABAA)受体α1(GABRA1)、β2(GABRB2)和γ2(GABRG2)亚基的mRNA和蛋白表达。结果与对照组相比,模型组大鼠精神状态较差,昼夜节律消失,体重、NREM睡眠和REM睡眠总量、下丘脑5-HT、GABA含量、GAD67阳性表达以及GABRA1、GABRB2、GABRG2的mRNA和蛋白水平显著降低,Wake总量、下丘脑Glu含量和Glu/GABA比值显著增加(P<0.05);与模型组相比,柏子养心汤低剂量、中剂量、高剂量组上述指标明显改善,且呈剂量依赖性。结论柏子养心汤可增加失眠大鼠NREM睡眠和REM睡眠总量,延长睡眠时间,改善睡眠质量,其作用机制可能与促进下丘脑GABA的合成,提高GABAA受体α1、β2和γ2亚基的表达有关。Objective To explore the influence of Baizi Yangxin Decoction on sleep phase in rats with insomnia induced by chlorophenylalanine(PCPA)based onγ-aminobutyric acid(GABA)signaling pathway.Methods SD rats were divided into control group,model group,and Baizi Yangxin decoction low-dose,medium-dose and high-dose groups(5.625 g/kg,11.25 g/kg,22.5 g/kg),with 24 rats in each group.A rat model of insomnia was established by intraperitoneal injection of PCPA.After 7 days of continuous gavage with corresponding doses of drugs,the general activity status and body weight changes of the rats were observed,electroencephalogram(EEG)and electromyography(EMG)signals were recorded by the animal sleep bioanalytical system,the total amount of wakefulness(Wake)and the total amount of non-rapid eye movement(NREM)and rapid eye movement(REM)sleep in rats at night and day were analyzed;the contents of 5-hydroxytryptamine(5-HT),glutamate(Glu)and GABA in the hypothalamus of rats were measured by enzyme-linked immunosorbent assay(ELISA);the expression of glutamate decarboxylase 67(GAD67)in the hypothalamus of rats was detected by immunohistochemistry(IHC);the mRNA and protein expressions ofγ-aminobutyric acid type A(GABAA)receptorα1(GABRA1),β2(GABRB2)andγ2(GABRG2)subunits in rat hypothalamus were measured by Western blot and RT-qPCR.Results Compared with control group,the rats in model group were in poorer mental state and the circadian rhythm disappeared,the body weight,total NREM and REM sleep,hypothalamic 5-HT,GABA content in the hypothalamus,GAD67 positive expression,and the mRNA and protein levels of GABRA1,GABRB2,and GABRG2 decreased obviously(P<0.05),while the total Wake,Glu content in hypothalamus and Glu/GABA ratio increased obviously(P<0.05);compared with model group,the above indexes were significantly improved in the low-,medium-and high-dose groups of Baizi Yangxin Decoction,in a dose-dependent manner.Conclusion Baizi Yangxin Decoction can increase the total NREM and REM sleep in insomnia rats,prolong sleep time,and improve

关 键 词：柏子养心汤 失眠 睡眠时相 Γ-氨基丁酸 Γ-氨基丁酸A受体 

分 类 号：R285.5[医药卫生—中药学]