常用失眠动物模型及其睡眠结构特征研究进展

失眠症是最为常见的睡眠障碍,以入睡和睡眠维持困难及日间功能障碍为主要表现。失眠症的病理生理研究和镇静催眠药物的非临床评价需以适当的动物模型为基础。本文以模型动物睡眠结构为重点,从心理应激、特殊睡眠环境和药物模型3个方面归纳常用失眠模型。现有失眠动物模型存在一定的局限性,在镇静催眠药物研究中应当灵活选择多种造模方法,以用更接近失眠临床表现的动物模型评价药物作用。     

四个新的失眠动物模型概述

失眠是一种影响健康的最常见疾病，是对睡眠时间和（或）质量不满足并影响白天社会功能的一种主观体验。失眠的发病率很高，我国成人失眠患病率高达57％，约50％的失眠症患者伴有各种精神疾病，因睡眠障碍引起的疾病占各类疾病总数的80％以上。失眠的形式有：睡眠潜伏期延长，睡眠维持障碍，睡眠质量下降，总睡眠时间缩短，日间残留头昏、精神不振、嗜睡、乏力等效应。按发生时段可分为起始失眠、间断失眠及终点失眠。临床常见的失眠有抑郁性失眠、焦虑性失眠、多梦性失眠、浅睡性失眠等类型。长期失眠会给正常生活和工作带来严重的不利影响。镇静催眠药物是治疗失眠的主要方法，失眠的治疗虽不复杂，但要治愈亦并不容易，由于临床失眠形式与类型的多样性，也给研究具有针对性的抗失眠药带来难度，新药研究仍然是一个值得重视的课题。目前国内的催眠药研究多数还未能应用失眠动物模型。因此，建立符合病因与发病机理的失眠动物模型以提高新药筛选与研究水平具有重要意义。本文简要介绍近年来几种新模型的制作方法，以供药物研究时参考选用。     

应激在抑郁症失眠中的作用和常用应激动物模型的睡眠特点

抑郁症导致的失眠常表现为睡眠中断和早醒, 慢波睡眠 (slow wave sleep, SWS) 减少或消失以及快速动眼 (rapid eye movement, REM) 睡眠潜伏期缩短。这些异常特征与应激引起的下丘脑-垂体-肾上腺 (hypothalamic-pituitary-adrenal, HPA) 轴去抑制亢进导致的睡眠异常极为相似, 提示两者之间可能有密切的联系。因此由应激引起的动物模型, 如创伤后应激障碍和慢性温和不可预知应激等模型, 可用来评价抗抑郁和改善睡眠的药物, 并为深入研究抑郁及伴发的失眠机制提供比较可靠的平台。本文重点阐述抑郁症失眠的特点、可能的病理生理机制、常用应激动物模型的建立方法并分析其睡眠结构。     

开心散对失眠模型大鼠睡眠周期的影响

摘要目的探讨开心散对失眠模型大鼠睡眠周期的影响。方法采用间断电流刺激法建立失眠大鼠模型。将48只大鼠随机分为6组，每组8只，分别为空白对照组，失眠对照组，地西泮（4 mg•kg－1）对照组，开心散低、中、高剂量组（2，4，8 g•kg－1），连续灌胃7 d后，通过皮层脑电描计方法，观察开心散对失眠大鼠睡眠周期的影响。结果失眠对照组觉醒时间（277.6±19.3） min，总睡眠时间（202.4±23.5） min；开心散中、高剂量组大鼠觉醒时间分别为（229.6±11.5），（211.4±13.1） min，睡眠总时间分别为（250.4±32.6），（268.6±29.7） min，且以慢波睡眠Ⅱ期（SWS2）和快动眼睡眠（REMS）的延长更显著（P＜0.05），其作用优于地西泮对照组。结论开心散具有显著的促睡眠效应，主要通过延长SWS2和REMS期来实现。     

失眠动物模型研究进展

催眠药物是对失眠症进行对因和对症治疗的首选药物。设计和选择良好的动物模型是催眠药物从临床前基础研究过渡到临床研究的关键环节之一。该文从模型制作的不同理论根据(中医理论和西医理论)出发,对近年来催眠药物研究所用的失眠动物模型的造模机制、具体方法、优缺点及适用情况作一简要综述。     

氟桂利嗪对戊巴比妥钠睡眠的作用及可能的作用机制

目的：探讨T型钙通道阻断剂氟桂利嗪对戊巴比妥钠睡眠的影响及可能的作用机制。方法：采用小鼠翻正反射试验及大鼠脑电（EEG）分析方法。结果：氟桂利嗪可增加大鼠总睡眠时间，非快眼动睡眠（NREMs）和快眼动睡眠（REMs）时间，其中主要延长深睡眠（SWS）时间，而对浅睡眠（Light）时间没有影响。还可剂量依赖性地延长阈上剂量戊巴比妥钠（45mg／kg，i．p．）诱导的小鼠睡眠时间并增加阈下剂量戊巴比妥钠（28mg／kg．i．p．）诱导的小鼠入睡率，     

失眠机制及药物治疗

随着工业化进程的加快,社会竞争日益加剧、工作压力、人口老龄化等原因,全球三分之一的人有睡眠问题,5%的人依靠药物维持着低质量的睡眠。临床使用的苯二氮卓艹类药物虽能增加睡眠量,但改变了睡眠脑电活动的生理模式,副作用多且易耐受。新型镇静催眠药主要包括:非苯二氮卓艹类、褪黑素受体激动剂、抗组胺药和食欲肽受体拮抗剂。这些药物起效快、疗效明显、"宿睡作用"少、耐受性及依赖性较低,已逐渐成为治疗失眠的主要手段。生理性睡眠调节异常是失眠的重要原因。由于缺乏失眠动物模型,限制了治疗药物开发。近年来,我们从生理性睡眠调节机制出发,探索能模拟人病理生理学特征的失眠动物模型,建立了紧张应激型、内稳态失衡型、生物钟紊乱型、神经网络功能异常、第一晚效应等失眠动物模型,用于镇静催眠药的筛选。中药是祖国医学的精华之一,但由于疗效观察多凭主观问卷,缺乏客观标准,加上中药成分复杂、作用机制不明、缺乏可靠的质控标准、使用不便等原因,限制其广泛应用和推广。我们利用高度自动化睡眠记录与解析平台,突破制约中药临床前疗效评价的瓶颈,运用现代药理学及睡眠神经生物学等研究手段,对可能具有镇静催眠效应的传统中药进行筛选和再评价,找出具有较强生理性睡眠调节作用的有效单体;利用基因敲除动物及各种失眠动物模型,结合神经化学、分子生物学、药理学等方法,从基因到行为研究这些有效单体的作用机制。结果发现,厚朴酚、厚朴酚、西红花素、西红花醛和白芍苷等能增加睡眠量,缩短睡眠潜伏期。脑电能谱分析显示,这些成分诱导的睡眠与生理性睡眠相似,提示这些成分可能适用于临床失眠症的治疗。     

生、酒五味子对失眠小鼠神经-内分泌-免疫网络的影响及机制

目的 探讨生、酒五味子对失眠小鼠神经-免疫-内分泌网络的影响及作用机制。方法 50只小鼠随机分为空白组、模型组、地西泮组、生五味子组、酒五味子组,每组10只。除空白组外,其余组小鼠腹腔注射甲状腺素溶液建立失眠小鼠模型,每天造模结束后分别灌胃相应剂量的地西泮和生、酒五味子。空白组和模型组给予等体积的生理盐水。观察并记录小鼠的一般状态;记录小鼠总活动距离及直立次数;记录小鼠的脑电和肌电信号,分析睡眠觉醒期（wake）、非快速眼动睡眠期（NREM）和快速眼动睡眠期（REM）所占时间比;检测小鼠脑视交叉上核（SCN）神经递质[γ-氨基丁酸（GABA）、5-羟色胺（5-HT）、多巴胺（DA）、去甲肾上腺素（NE）、皮质醇（CORT）]含量,检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)的荧光强度;检测时钟基因Bmal1、生物钟基因Clock、周期基因Per2 mRNA的表达。结果 与空白组比较,模型组小鼠精神状态相对萎靡,饮食量和饮水量增加,体质量降低,毛发粗糙无光泽,昼夜节律出现不规律现象;总活动距离、直立次数显著降低;wake所占时间比显著上升,REM、NREM所占时间比...     

24 h 快速眼动睡眠剥夺大鼠血清的蛋白质组学初步研究

摘要： 目的 探讨睡眠剥夺大鼠血清蛋白质组中差异表达蛋白质及其意义。方法 采用改良多平台水环境法 建立 24 h 快速眼动睡眠剥夺大鼠模型。采用随机数字表法将大鼠均分为模型（M）组、 模型对照（MC）组、 空白对照 （BC） 组, 每组 8 只大鼠。观察大鼠体质量变化, 采用 Morris 水迷宫实验检测睡眠剥夺对大鼠学习记忆能力的影响, 并用双向电泳技术筛选血清差异蛋白, LC-MS/MS 技术鉴定差异蛋白。结果 BC 组和 MC 组大鼠建模前后体质量 差异均无统计学意义。M 组大鼠毛发光泽下降, 精神稍差, 较亢奋, 体质量在睡眠剥夺后显著降低。24 h 快速眼动 睡眠剥夺对大鼠学习记忆能力无显著性影响。各组之间蛋白质点数目差异无统计学意义。质谱鉴定出 4 种表达下 降的蛋白, 分别为 Serotransferrin; Glutathione peroxidase 3; Ig kappa chain C region, B allele; Collagen alpha-2(I) chain。 结论 短期睡眠剥夺对机体的损伤可能与铁离子代谢、 氧化应激和免疫功能等有关     

小剂量米安色林治疗慢性失眠初期效果评价

目的：评价小剂量米安色林（7.5 mg）治疗慢性失眠初期效果。方法：选择慢性失眠非严重期患者20例，米安色林7.5 mg·d^-1，研究周期为21 d，在第2~3周期间停药2~3 d后安慰剂治疗1 d。用匹兹堡睡眠质量指数量表（PSQI）评分对患者治疗后1周、第3周与治疗前对比评定疗效，副反应量表（TESS）评定药物不良反应。第8~20天多导睡眠监测（PSG）评估药物和安慰剂治疗期睡眠潜伏期（SL）、总睡眠时间（TST）、快速眼动睡眠时间（REM）、慢波睡眠时间（N₃）的变化，比较药物与安慰剂疗效差异。结果：治疗3周PSQI评分减少到（3.9±2.2），与治疗前评分（14.3±2.1）对比有显著性差异（P<0.05），PSG结果显示药物较安慰剂治疗增加了TST和N₃期时间，2组对比（435.5±45.2/411.2±71.6，16.5±15.6/9.6±10.0），统计学分析有显著性差异（P<0.05），结果证实小剂量米安色林治疗失眠增加睡眠时间和深睡眠时间效果显著。2例出现有轻度头晕和思睡，无需要特殊处理。结论：小剂量米安色林治疗慢性失眠初期有效，增加睡眠时间和深睡眠时间效果显著，有改善睡眠效应。     

Effects of short-acting hypnotics on sleep latency in rats placed on grid suspended over water

The present study was performed to develop a new sleep disturbance model for evaluating hypnotic potencies by placing rats on a grid suspended over water up to 1 cm under the grid surface. When rats were placed on the grid, significant increases in sleep latency and amount of wakefulness were observed compared with those of rats placed on sawdust. However, the amounts of non-rapid eye movement (non-REM) sleep and rapid eye movement (REM) sleep of rats placed on the grid were significantly decreased compared with those of rats placed on sawdust. Four short-acting hypnotics (triazolam, zopiclone, brotizolam, lormetazepam) caused significant decreases in sleep latency, and the effects of hypnotics in rats placed on the grid were more potent than those in rats placed on sawdust. In conclusion, the present model can serve as a new sleep disturbance model and may also be useful for evaluating the sleep-inducing effects of short-acting hypnotics.     

Effects of SDZ NVI-085, a putative subtype-selective alpha 1-agonist, on canine cataplexy, a disorder of rapid eye movement sleep.

Canine narcolepsy is an animal model of the human rapid eye movement sleep disorder. Dogs exhibit bouts of sleep attacks and muscle atonia (cataplexy) that are induced by emotions and thought to be abnormal rapid eye movement sleep episodes. We have previously demonstrated that cataplexy is strongly inhibited by increases in noradrenergic activity. This effect is mediated through central alpha 1-adrenoceptors, presumably of the alpha 1B subtype. In this study, we demonstrate with the canine model that SDZ NVI-085, a new compound with alerting effects, is a potent anticataplectic agent that may act through stimulation of an alpha 1-adrenoceptor subtype.     

Oleamide restores sleep in adult rats that were subjected to maternal separation

Maternal separation (MS) induces a series of changes in rats' behavior; among them a reduction in spontaneous sleep. One potentially impaired system is the endocannabinoid system (eCBs), since it contributes to generate sleep. To investigate if there are situations early in life that affect the eCBs, which would contribute to make rats vulnerable to suffering insomnia, we studied the rodent model of MS. Rats were separated from their mothers for 3 h-periods daily, from postnatal day (PND) 2 to PND 16. Once they gained 250 g of body weight (adult rats), they were implanted with electrodes to record the sleep-waking cycle (SWC). MS rats and non-MS (NMS) siblings were assigned to one of the following groups: vehicle, oleamide (OLE, an agonist of the cannabinoid receptor 1, CB1R), OLE + AM251 (an antagonist of the CB1R) and AM251 alone. Expression of the CBR1 receptor was also analyzed in the frontal cortex (FCx) and in the hippocampus (HIP) of both NMS and MS rats. Results indicated that MS induced a reduction in both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep with the consequent increase in waking (W) as compared to NMS siblings. OLE normalized the SWC, and AM251 blocked such an effect. CB1R expression was reduced in the FCx and in the HIP of MS rats. Our results indicate that MS reduces sleep and CB1R expression and OLE improves sleep in adult rats.     

安定类药物对失眠症患者日间功能和睡眠质量的影响

目的探讨长期使用安定类药物对失眠症患者日间功能和睡眠质量的影响。方法对35例长期使用安定类药物的老年失眠症患者（研究组）进行整夜多导睡眠图描记和日间功能问卷调查,并与30例未使用安定类药物的老年失眠症患者（对照组）对照分析。结果研究组和对照组日间功能异常者分别为91.4%和30.0%,两组差异有统计学意义（P〈0.05）;多导睡眠图显示研究组睡眠潜伏期缩短、睡眠时间延长、浅睡眠增多、深睡眠和快速眼动期（REM）睡眠减少,与对照组比较差异有统计学意义（P〈0.01）。结论长期使用安定类药物可导致睡眠质量和日间功能下降。     

Effects of kava-kava extract on the sleep–wake cycle in sleep-disturbed rats

Rationale Kava-kava extract may be useful as an herbal medicine for treatment of insomnia and anxiety.Objectives The present study was undertaken to investigate the effects of kava-kava extract on the sleep–wake cycle in comparison with that of flunitrazepam using sleep-disturbed rats.Methods Electrodes for measurement of electroencephalogram (EEG) and electromyogram (EMG) were implanted into the frontal cortex and the dorsal neck muscle of rats. EEG and EMG were recorded with an electroencephalogram. SleepSign ver.2.0 was used for EEG and EMG analysis. Total times of wakefulness, non-rapid eye movement (non-REM) and REM sleep were measured from 09:00 to 15:00.Results A significant shortening of the sleep latency in sleep-disturbed rats was observed following the administration of kava-kava extract at a dose of 300 mg/kg, while no effects were observed on the total waking and non-REM sleep time. On the other hand, flunitrazepam showed a significant shortening in sleep latency, decrease in total waking time and increase in total non-REM sleep time. Although the effects of flunitrazepam were antagonized by the benzodiazepine receptor antagonist flumazenil, the effect of kava-kava extract was not antagonized by flumazenil. Kava-kava extract showed a significant increase in delta activity during non-REM sleep in sleep-disturbed rats, whereas a significant decrease in delta power during non-REM sleep was observed with flunitrazepam. Flumazenil caused no significant effect on the changes in delta activity induced by both kava-kava extract and flunitrazepam.Conclusions Kava-kava extract is an herbal medicine having not only hypnotic effects, but also sleep quality-enhancement effects.     

Effect of tandospirone on sleep latency in rats placed on a grid suspended over water

The present study was performed to examine the effect of tandospirone on sleep latency in a new insomnia animal model by placing rats on a grid suspended over water. For investigating the mechanism of tandospirone, the effect of tandospirone on sleep latency was also studied using rats that were depleted with neuronal serotonin (5-HT) after p-chlorophenylalanine administration. Tandospirone caused a shortening of sleep latency dose-dependently, and a significant effect was observed at 20 mg/kg, p.o. or more. A shortening of sleep latency was observed by administration of p-chlorophenylalanine (300 mg/kg, i.p.) for 2 days. On the other hand, tandospirone exerted no potentiating effect on the shortening of sleep latency induced by p-chlorophenylalanine. From these findings, a shortening of sleep latency induced by tandospirone may occur through the pre-synaptic 5-HT(1A) receptors in rats.     

Effects of some H1-antagonists on the sleep-wake cycle in sleep-disturbed rats

The present study was undertaken to investigate the effects of some H(1)-antagonists on the sleep-wake cycle in sleep-disturbed rats in comparison with those of nitrazepam. Electrodes were chronically implanted into the frontal cortex and the dorsal neck muscle of rats for the electroencephalogram (EEG) and electromyogram (EMG), respectively. EEG and EMG were recorded with an electroencephalograph. SleepSign ver. 2.0 was used for EEG and EMG analysis. The total times of waking, non-rapid eye movement (non-REM), and rapid eye movement (REM) sleep were measured from 10:00 to 16:00. Nitrazepam showed a significant decrease in sleep latency, total waking time, and delta activity and an increase in the total non-REM sleep time. A significant decrease in the sleep latency was observed with diphenhydramine, chlorpheniramine, and cyproheptadine. Cyproheptadine also caused a significant decrease in the total waking time and increases in total non-REM sleep time, REM sleep time, slow wave sleep, and delta activity, although no remarkable effects were observed with diphenhydramine and chlorpheniramine. In conclusion, cyproheptadine can be useful as a hypnotic, having not only sleep inducing-effects, but also sleep quantity- and quality-increasing effects.     

小鼠睡眠生物解析系统与应用

目的建立高度自动化小鼠睡眠生物解析系统,并考察作用于多巴胺D1/D2受体的左旋千金藤啶碱(L-stepholi-dine,SPD)对小鼠睡眠的影响。方法动物饲养在恒温、恒湿、隔音、静电屏蔽、12h/12h明暗光照环境下,利用Sleep-Sign软件,连续48h记录自由运动小鼠的脑电和肌电,分别在d121:00点腹腔注射生理盐水和d2同一时间注射SPD;并用此软件对脑电和肌电进行分析,自动判断睡眠-觉醒时相。结果该系统能高度自动化和高效率地记录与解析小鼠睡眠-觉醒行为。小鼠觉醒期腹腔注射SPD能增加睡眠量,延长非快动眼睡眠波的时程,降低觉醒量及觉醒期脑电波的强度,但不影响快动眼睡眠。结论作用于多巴胺受体的SPD能促进非快动眼睡眠,抑制觉醒。     

Effect of cannabidiol on sleep disruption induced by the repeated combination tests consisting of open field and elevated plus-maze in rats

Patients with post-traumatic stress disorder (PTSD) frequently complain of having sleep disturbances, such as insomnia and rapid eye movement (REM) sleep abnormality. Cannabidiol (CBD), a psycho-inactive constituent of marijuana, reduces physiological non-REM (NREM) sleep and REM sleep in normal rats, in addition to generating its anxiolytic effect. However, the effects of CBD on anxiety-induced sleep disturbances remain unclear. Because anxiety progression is caused by persistent stress for a period of time, we employed the repeated combination tests (RCT) consisting of a 50-min open field (OF) and a subsequent 10-min elevated plus-maze (EPM) for four consecutive days to simulate the development of anxiety. Time spent in the centre arena of OF and during open arms of the EPM was substantially decreased in latter days of RCT, suggesting the habituation, which potentially lessens anxiety-mediated behavioural responses, was not observed in current tests. CBD microinjected into the central nucleus of amygdala (CeA) significantly enhanced time spent in centre arena of OF, increased time during the open arms and decreased frequency of entry to the enclosed arms of EPM, further confirming its anxiolytic effect. The decrease of NREM sleep during the first hour and the suppression of REM sleep during hours 4-10 after the RCT represent the similar clinical observations (e.g. insomnia and REM sleep interruption) in PTSD patients. CBD efficiently blocked anxiety-induced REM sleep suppression, but had little effect on the alteration of NREM sleep. Conclusively, CBD may block anxiety-induced REM sleep alteration via its anxiolytic effect, rather than via sleep regulation per se. This article is part of a Special Issue entitled 'Anxiety and Depression'.