快速眼球运动睡眠剥夺及γ-氨基丁酸能药物干预对大鼠认知功能的影响  

Effects of rapid eye movement sleep deprivation and GABAergic drug intervention on cognition in rats

作　　者：李振[1] 赵忠新[1] 黄流清[1] 向正华[2] 张琳[1] 张鹏[1] 李雁鹏[1] 吴惠涓[1] 庄建华[1] 王文昭[1] 贺斌[1] 

机构地区：[1]第二军医大学附属长征医院神经内科,上海200003 [2]第二军医大学神经科学研究所

出　　处：《中华神经科杂志》2011年第8期538-543,共6页Chinese Journal of Neurology

基　　金：国家自然科学基金资助项目（30900473,81070070）;军队特需药品研究技术平台建设基金资助项目（2011ZXJ092301）;上海市科技发展基金资助项目（08411950700）

摘　　要：目的建立快速眼球运动（REM）睡眠剥夺和下丘脑穹隆周围核微量渗析大鼠模型，研究不同程度REM睡眠剥夺和恢复对SD大鼠认知功能、下丘脑泌素（Hcrt）能系统和γ-氨基丁酸（GABA）能系统的影响及其可能机制。方法将雄性SD大鼠随机分为对照组和REM睡眠剥夺组，REM睡眠剥夺组又分非手术对照组（nonOP）、假手术对照组（Sham）、GABA。受体拮抗剂SR-95531组（SR）和GABA再摄取抑制剂NO-711组（NO）。采用改良多平台水环境法（MMPM）建立REM睡眠剥夺SD大鼠模型，利用Morris水迷宫测定大鼠在不同程度REM睡眠剥夺对认知功能的影响变化，采用免疫荧光技术检测下丘脑Herr神经元数量形态和原癌基因（Fos）表达、下丘脑GABA的A型受体α1（GABAARα1）亚单位表达累积吸光度值；采用高效液相色谱仪（HPLC）测定下丘脑GABA和谷氨酸（Glu）含量。建立穹隆周围核微量渗析SD大鼠模型，观察GABAA受体拮抗剂SR-95531和GABA再摄取抑制剂NO-711对上述指标的影响。结果REM睡眠剥夺导致nonOP组和Sham组SD大鼠认知功能下降、下丘脑外侧区Fos阳性细胞总数和Fos—Hcrt双阳性细胞（F^＋＆H^＋）数量增加、下丘脑GABA含量和GABAARα1表达增加，且变化程度均与REM睡眠剥夺时间长短呈正相关。但上述指标的变化都能够在不同程度睡眠恢复后得以修正。NO组与对照组比较，睡眠剥夺期间认知功能下降、下丘脑外侧区Fos阳性细胞总数和F^＋＆H^＋数量增加（F=9．56、12．14、10．12，P〈0．05），且增加程度均与REM睡眠剥夺时间长短呈正相关；但睡眠恢复后差异无统计学意义。SR组与对照组比较，睡眠剥夺期间差异无统计学意义，但睡眠恢复期间认知功能下降、下丘脑外侧区Fos阳性细胞总数和F^＋＆H^＋数量增加（F=12．03、11．38、8．36，均P〈0．05）。SR组GABA含量和GABAARα1表达在全部5个时间点均明显�Objective To establish an animal model of rapid eye movement （REM） sleep deprivation （SD） and an animal model for perifornical nucleus microdialysis and investigate the change of cognition, hypocretinergic system and GABAergic system in rats＇ hypothalamus after various degrees of REM sleep deprivation and sleep revival and two GABAergic drugs intervention. Methods The modified multiple platform method （MMPM）was used to establish sleep deprivation model and the cognitive function was assessed by Morris＇ water maze. Immunofluorescence technique was used to analyze the number of Hypocretin （Hcrt） immunoreactive neurons, total Fos immunoreactive neurons, Hcrt and Fos colabeled neurons, and the integrated optical density （IA） of GABAARα1 immunoreactive area in rats＇ hypothalamus. High performance liquid chromatograph （HPLC） was used to quantitatively analyze the level of GABA and Gluin in the rats＇ hypothalamus. Two GABAergic drugs, a selective GABAAR antagonist, SR-95531, and a selective blocker of type 1 GABA transporter （uptake blocker）, NO-711, were used for perifornical nucleus microdialysis. Results There was no statistically significant difference in tests between CC and TC （Define CC and TC）. There was a significant decrease （P 〈 0. 05 ） of cognitive function measured by Morris maze test in SD 3 d, SD 5 d and RS 6 h of SD groups compared with CC and TC groups. Number of Fos immunoreactive, F^＋ ＆H ^＋ immunoreactive neuronsand IA of GABAA Roll immunoreactive area were all significantly increased （P 〈 0. 05 ）. Content of GABA measured by HPLC was also increased （ P 〈 0. 05 ）. However, all these changes were partly reversed by sleep revival. SR-95531 and NO-711 had different effect on these changes. Conclusions Sleep deprivation, no matter mild or severe, has adverse effects on cognitive function. Activities of both GABAergic and Hcrtergic system are increased during REMSD. These two neurons system could be regulated by each other and the relationshi

关 键 词：睡眠剥夺 认知 Γ-氨基丁酸 神经肽类 细胞内信号肽和蛋白质类 大鼠 

分 类 号：R740[医药卫生—神经病学与精神病学]