丘脑室旁核腺苷A1受体表达变化及其在睡眠调控中的作用  

Altered expression of adenosine A1 receptor in the paraventricular thalamus and its role in the regulation of sleep

作　　者：朱一峰 王娜[1] 胡俊雅 雷梦珠 夏建霞[1] 王亚玲[1] 胡志安[1] ZHU Yifeng;WANG Na;HU Junya;LEI Mengzhu;XIA Jianxia;WANG Yaling;HU Zhi’an(Department of Physiology,College of Basic Medical Sciences,Army Medical University(Third Military Medical University),Chongqing;Chongqing Institute for Brain and Intelligence,Chongqing;Department of Sleep and Medical Psychology,Chongqing Health Center for Women and Children,Chongqing,China)

机构地区：[1]陆军军医大学(第三军医大学)基础医学院生理学教研室,重庆 [2]重庆脑与智能科学中心,重庆 [3]重庆市妇幼保健院睡眠心理科,重庆

出　　处：《陆军军医大学学报》2025年第8期766-774,共9页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(CSTB2024NSCQ-MSX1007);重庆市自然科学基金创新发展联合基金(CSTB2024NSCQ-LZX0113)。

摘　　要：目的 探究丘脑室旁核(paraventricular thalamus,PVT)内腺苷A1受体在觉醒/睡眠状态下的表达模式及其在调控睡眠中的作用。方法 选取6~12周龄雄性C57BL/6J小鼠(体质量22~28 g)为实验动物,按照随机数字表法将实验动物分为ZT0组、ZT6组、ZT10组、ZT12组、睡眠剥夺组和恢复睡眠组(n=16或n=4),采用RT-qPCR和荧光原位杂交方法,检测PVT脑区腺苷A1受体RNA表达变化。采用离体全细胞膜片钳记录方法,观察腺苷对小鼠PVT神经元膜电位和放电频率的影响。将实验动物按照随机数字表法分为腺苷A1受体干扰组(n=8)和干扰对照组(n=7),采用RNA干扰和脑肌电记录技术,观察在睡眠剥夺6 h、恢复睡眠4 h过程中2组觉醒及睡眠时间变化。结果 RT-qPCR和荧光原位杂交实验表明小鼠PVT中腺苷A1受体受觉醒/睡眠状态的影响,ZT0组(活跃期)表达明显高于ZT12组(非活跃期,P<0.05)。全细胞膜片钳结果显示腺苷对PVT神经元的抑制效应有2种反应类型,随腺苷A1受体表达水平降低而减弱(P<0.05)。睡眠剥夺后,睡眠压力越大,腺苷A1受体表达水平呈现显著组间差异:睡眠剥夺组显著高于恢复睡眠组(P<0.05),而恢复睡眠组又高于正常昼夜节律的ZT6组和ZT10组(P<0.05)。与干扰对照组相比,PVT脑区腺苷A1受体干扰组在恢复睡眠1 h内觉醒时间明显升高,睡眠时间明显下降(P<0.05)。结论 PVT内腺苷A1受体的表达受到睡眠压力的动态调控,表达水平随着睡眠压力增大而升高。Objective To explore the expression profile of adenosine A1 receptor in the paraventricular thalamic nucleus(PVT)in wakefulness/sleep state and its role in regulating sleep.Methods Male C57BL/6J mice(6~12 weeks old,weighing 22~28 g)were randomly divided into ZT0,ZT6,ZT10,ZT12,and sleep deprivation and recover0y sleep groups(n=16 or n=4).RT-qPCR and fluorescence in situ hybridization were used to observe the changes of adenosine A1 receptor in PVT.Whole-cell patch-clamp recording was conducted to determine the effects of adenosine on the membrane potential and discharge frequency of PVT neurons.The experimental animals were divided into adenosine A1 receptor interference group(n=8)and interference control group(n=7).After RNA interference,electroencephalography(EEG)and electromyography(EMG)were applied simultaneously to observe the changes in wake and sleep time between the 2 groups during sleep deprivation for 6 h and sleep recovery for 4 h.Results RT-qPCR and fluorescence in situ hybridization confirmed that the expression of adenosine A1 receptor in PVT was affected by wakefulness/sleep state,with the level in the ZT0 group(active stage)significantly higher than that in the ZT12 group(non-active stage,P<0.05).Whole-cell patch clamp recording indicated that adenosine exerted inhibitory effects on PVT neurons through 2 distinct response types(P<0.05),and the inhibition was in a diminishing trend with a decrease in the expression level of adenosine A1 receptor.After sleep deprivation,the expression level of adenosine A1 receptor showed significant intergroup differences:the level was significantly higher in the sleep deprivation group than the recovery sleep group(P<0.05),while that of the recovery sleep group was higher than that of the ZT6 group and that of the ZT10 group(P<0.05).After knocking down adenosine A1 receptor(shRNA-A1R)in the PVT,the wakefulness timing of the shRNA-A1R group was significantly increased,while the sleep timing was significantly decreased within 1 h after sleep recovery when compared wi

关 键 词：腺苷A1型受体 丘脑室旁核 觉醒/睡眠 睡眠压力 

分 类 号：R338.24[医药卫生—人体生理学] R338.63[医药卫生—基础医学] R341