5-HT_(1A)和5-HT_2受体功能与觉醒、睡眠成分关系的研究

目的　观察 5 HT1A受体激动剂 8 OH DPAT和HT2受体拮抗剂利坦色林 (ritanserin)对大鼠清醒和睡眠成分的影响 ,给予 5 HT1A受体激动剂和剥夺REM睡眠后皮层 5 HT2 受体结合能力的变化 ,进一步分析两种 5 HT受体亚型之间的关系以及 5 HT2 受体在睡眠中的作用。方法　利用大鼠睡眠自动分析系统定量分析清醒和睡眠成分的变化 ,注射PCPA制作剥夺睡眠的大鼠模型 ,水上平台法制作剥夺REM睡眠的大鼠模型 ,利用放射配体结合实验研究配体与受体结合力的变化。结果　 8 OH DPAT小剂量 (0 0 1mg·kg-1,sc)可以增加深睡眠和浅睡眠 ,减少觉醒 ;大剂量(0 375mg·kg-1,sc)则增加觉醒 ,减少全部睡眠成分。 5 HT2 受体拮抗剂ritanserin可以明显增加深睡眠 ,减少觉醒和REM睡眠。而 8 OH DPAT低剂量与ritanserin联合用药则使深浅睡眠明显增加 ,清醒和REM睡眠明显减少 ,具有协同作用。对于PCPA化使 3种睡眠成分均明显减少 ,觉醒比例明显增加的大鼠 ,ritanserin仅使浅睡眠增加其它成分不变。用 [3 H] ritanserin放射性配基结合显示 ,剥夺大鼠REM睡眠后皮层 5 HT2 受体Bmax值明显增加但Kd 值无变化 ;给予 5 HT1A激动剂 8 OH DPAT后皮层 5 HT2 受体Kd 和Bmax均降低。结论　 5 HT1A受体和 5 HT2 受体与睡眠有密切关系     

中缝背核投射到基底外侧杏仁核的5-羟色胺能纤维对睡眠的调节作用

目的　观察大鼠中缝背核 (DRN)到基底外侧杏仁核(BLA)的 5 HT能纤维投射在睡眠 -觉醒调节中的作用。方法　采用脑立体定位 ,核团微量注射和多导睡眠描记 (PSG)方法。结果　DRN内微量注射L Glu ,可使觉醒 (W )增加 ,慢波睡眠 (SWS)和异相睡眠 (PS)明显减少。在双侧BLA微量注射非选择性 5 HT受体阻断剂麦角新碱 (MS)可以逆转DRN内微量注射L Glu的效应 ,SWS增加 ,W减少 ,但PS没有变化 ;DRN内微量注射PCPA ,导致SWS增加 ,W减少 ,但当在DRN内微量注射PCPA后 ,双侧BLA内微量注射 5 HTP可以逆转PCPA所引起的睡眠增加效应 ,使SWS减少 ,W增加 ,但PS没有变化。结论　DRN对睡眠 -觉醒的调节作用部分通过DRN到BLA的 5 HT能纤维投射介导的     

腹外侧视前区5-羟色胺对大鼠睡眠-觉醒周期的作用

目的　通过腹外侧视前区 (VLPO)微量注射 5 羟色胺酸 (5 HTP)、非特异性 5 HT受体阻断剂麦角新碱 (MS)和5 HT再摄取抑制剂氟西汀观察 5 HT在VLPO对大鼠睡眠-觉醒周期的影响。方法　采用脑立体定位、核团微量注射和多导睡眠描记技术。结果　VLPO双侧分别微量注射小剂量 5 HTP(0 5μg ,0 1 μl)对大鼠睡眠 -觉醒周期无明显影响 ;而双侧注射大剂量 5 HTP(1 0 μg ,0 1 μl)使大鼠睡眠减少 ,觉醒增加 ;VLPO双侧注射 5 HT再摄取抑制剂氟西汀(6 0 μg ,0 1 μl)可产生与大剂量 5 HTP类似的作用 ;而VLPO双侧微量注射非特异性 5 HT受体阻断剂麦角新碱(MS ,1 0 μg ,0 1 μl)使大鼠睡眠增加 ,觉醒减少。大剂量 5 HTP和MS对觉醒 ,睡眠成分改变具有明显的时间相关性。结论　 5 HT在VLPO参与睡眠 -觉醒周期调节且有促觉醒作用 ,其促觉醒作用可能与突触后物质表达有关     

5-HT_(1A)受体激动剂乌拉地尔对吗啡依赖大鼠前列腺组织的病理学影响

目的··:观察5-HT1A 受体激动剂乌拉地尔对吗啡依赖大鼠前列腺的组织学影响。方法··:皮下注射(sc)5d吗啡 ,建立吗啡依赖大鼠模型 ;实验组分别进行侧脑室注射 (icv)乌拉地尔和继续sc吗啡。实验后将前列腺组织作HE染色后在光镜下观察。结果··:吗啡依赖大鼠的前列腺组织有轻度萎缩 ,吗啡依赖大鼠自然戒断后前列腺有明显增生 ,icv乌拉地尔可抑制吗啡戒断大鼠的前列腺组织增生。结论··:乌拉地尔可抑制吗啡戒断大鼠前列腺组织的增生     

5-HT_(1P)受体介导的交感神经节慢突触传递

目的　在豚鼠肠系膜下神经节 (IMG)细胞上观察不同 5 羟色胺 (5 HT)受体亚型拮抗剂对非胆碱能迟慢兴奋性突触后电位 (LS EPSP)的作用。方法　应用离体细胞内记录技术。结果　赛庚啶 (cyproheptadine ,5 HT1/ 2 受体拮抗剂 )及BRL 2 492 4(5 HT1P受体拮抗剂 )可逆的阻抑 5 HT敏感细胞的LS EPSP ,而mianserin(5 HT2 受体拮抗剂 ) ,MDL72 2 2 2 (5 HT3 受体拮抗剂 ) ,spiperone(5 HT1A受体拮抗剂 )对LS EPSP无显著影响 ;用MCPP(5 HT1P受体激动剂 )持续灌流IMG使 5 HT敏感细胞脱敏后可逆的阻抑LS EPSP的发生。结论　 5 HT敏感细胞的LS EPSP是由 5 HT1P受体介导的。     

5—HT2C受体：设计抗肥胖药物和抗癫痫药物的潜在治疗靶标

5－羟色胺（5－HT）是一种重要的神经递质，在哺乳动物的中枢神经系统中至少有14种不同的5－HT受体亚型，研究发现其中5－HT2C受体对食物摄取和癫症的发作有调节作用。5－HT2C受体激动剂可能是通过增加饱食感来减少对食物的提取，许多高选择性的5－HT2C受体激动剂正进行临床前及临床评价，5－HT2C受体有可能成为抗肥胖和抗癫痫药物的新靶标。     

孤束核微注射5-羟色胺系统药物对大鼠睡眠-觉醒的影响及机制

目的:探讨孤束核(NST)微注射5-羟色胺(5-HT)系统药物对大鼠睡眠-觉醒的影响及其机制.方法:选用雄性SD大鼠,分7组,孤束核分别微注射5-羟基色氨酸(5-HTP)、非特异性5-HT受体阻断剂麦角新碱(MS)、5-羟色胺1A受体(5-HT1AR)激动剂8-OH-DPAT、5-HT1AR拮抗剂spiperone、cAMP、cAMP依赖的蛋白激酶A(PKA)拮抗剂H89和生理盐水(NS)后,采用光电脑电机记录脑电和肌电;采用免疫组化ABC法检测NST处5-HT1AR及5-HT表达情况.结果:与处理前相比,NS组和MS组大鼠睡眠-觉醒各期差异均无统计学意义,5-HTP组、8-OH-DPAT组和H89组给药后均显示觉醒期(W)缩短,异相睡眠(PS)延长(P<0.05或P<0.01),8-OH-DPAT组和H89组慢波睡眠(SWS)延长(P<0.01或P<0.05);spiperone组和cAMP组W延长(P<0.01或P<0.05),SWS缩短(P<0.01或P<0.05),spiperone组PS缩短(P<0.05).免疫组化结果显示,与NS组相比,5-HTP组的5-H T1AR阳性细胞数较多,5-HT能神经纤维末梢5-HT表达增强,MS组则结果相反.结论:作为睡眠诱发区,孤束核很可能是通过5一羟色胺作用于5-HT1AR,使cAMP生成减少,PKA磷酸化减少而发挥使觉醒减少、睡眠增加的生物学效应.     

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

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...     

莫达非尼对REM睡眠剥夺后大鼠认知功能及突触可塑性的影响*

目的：探讨新型中枢性兴奋剂莫达非尼对不同程度快速动眼（REM）睡眠剥夺与睡眠恢复后大鼠认知功能和大鼠海马突触可塑性的影响。方法：成年雄性SD大鼠随机分为莫达非尼组与对照组,每组分5个不同的REM睡眠剥夺和恢复时间点（SD1d,SD3d,SD5d,SD5d/RS6h,SD5d/RS12h）,采用改良多平台水环境REM睡眠剥夺法进行REM睡眠剥夺,利用Y迷宫测定大鼠学习记忆能力,运用蛋白质免疫印记（Western Blot）技术对大鼠海马突触素（SynapsinⅠ）蛋白作选择性半定量分析,免疫组织化学和电镜观察的方法分析大鼠海马CA3区SynapsinⅠ的表达,观察突触可塑性变化。结果：莫达非尼组的错误反应次数（EN）在REM睡眠剥夺后小于对照组,差异有统计学意义（P〈0.05）,睡眠恢复后两组EN比较,差异无统计学意义（P〉0.05）,总反应时间（TRT）两组比较,差异也无统计学意义（P〉0.05）。Western Blot结果显示SD1d,SD3d,SD5d/RS6h和SD5d/RS12h时莫达非尼组Syn- apsinⅠ表达比对照组增多（P〈0.05）,SD5d时两组SynapsinⅠ表达无显著差异（P〉0.05）,免疫组化结果与其一致;电镜下观察SD1d时莫达非尼组的突触联系、突触前膜囊泡数量均较对照组增加。结论：REM睡眠剥夺会引起大鼠认知功能下降,大鼠海马CA3区SynapsinⅠ表达降低,而莫达非尼可以抑制睡眠剥夺后SynapsinⅠ的表达减少,诱导大鼠海马突触可塑性变化,可改善REM睡眠剥夺后的认知行为。     

阻塞性睡眠呼吸暂停低通气综合征与认知障碍

阻塞性睡眠呼吸暂停低通气综合征(obstructive sleep apnea-hypopnea syndrome,OSAHS)是一种严重的睡眠呼吸疾病,以反复发作的睡眠呼吸暂停和低氧血症为主要临床特征,常出现睡眠结构紊乱,表现为浅睡眠和觉醒增加,快速眼动睡眠(rapid eye movement sleep,REMS)和深睡眠时间减少,并引起白天困倦嗜睡。一般人群OSAHS的发病率为2%～5%,60     

前列腺素D2的睡眠调节作用

前列腺素D2(PGD2) 是由前列腺素D2合成酶合成的一种二十碳不饱和脂肪酸.研究表明PGD2具有睡眠调节作用,其机理为PGD2与前列腺素D2受体结合,升高基底前脑部位细胞外腺苷水平.腺苷通过腺苷A2A受体激活睡眠调节中枢(腹外侧视前区,VLPO)神经元,并通过GABA抑制觉醒调节中枢(结节乳头核,TMN)组胺能神经元而导致睡眠.对PGD2睡眠调节作用及其机理进行研究,有望开发出新型的镇静催眠药物.     

Discovery and development of orexin receptor antagonists as therapeutics for insomnia

Insomnia persistently affects the quality and quantity of sleep. Currently approved treatments for insomnia primarily target γ-aminobutyric acid-A (GABA-A) receptor signalling and include benzodiazepines and GABA-A receptor modulators. These drugs are used to address this sleep disorder, but have the potential for side effects such as tolerance and dependence, making them less attractive as maintenance therapy. Forward and reverse genetic approaches in animals have implicated orexin signalling (also referred to as hypocretin signalling) in the control of vigilance and sleep/wake states. Screening for orexin receptor antagonists using in vitro and in vivo methods in animals has identified compounds that block one or other of the orexin receptors (single or dual orexin receptor antagonists [SORAs and DORAs], respectively) in animals and humans. SORAs have primarily been used as probes to further elucidate the roles of the individual orexin receptors, while a number of DORAs have progressed to clinical development as pharmaceutical candidates for insomnia. The DORA almorexant demonstrated significant improvements in a number of clinically relevant sleep parameters in animal models and in patients with insomnia but its development was halted. SB-649868 and suvorexant have demonstrated efficacy and tolerability in Phase II and III trials respectively. Furthermore, suvorexant is currently under review by the Food and Drug Administration for the treatment of insomnia. Based on the publication of recent non-clinical and clinical data, orexin receptor antagonists potentially represent a targeted, effective and well-tolerated new class of medications for insomnia.Linked ArticlesThis article is part of a themed section on Orexin Receptors. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-2     

Effects of ramelteon 8 mg on objective sleep latency in adults with chronic insomnia on nights 1 and 2: pooled analysis*

ABSTRACT

Objective: Ramelteon is an MT₁/MT₂ melatonin receptor agonist indicated for the treatment of insomnia characterized by difficulty with sleep onset. In previous clinical studies, ramelteon reduced latency to persistent sleep (LPS) in subjects with chronic insomnia. The goal of the current analysis was to determine the average reduction in LPS and overall adverse event profile for subjects taking ramelteon 8?mg.

Research design and methods: This pooled analysis examined four randomized, double-blind, placebo-controlled clinical trials of ramelteon in subjects with chronic insomnia. The analysis included adults (age 18–83?years) with chronic insomnia who took ramelteon 8?mg or placebo. The primary endpoint of each trial was mean LPS, measured by polysomnography (PSG) on nights 1 and 2. Adverse events were collected for all subjects for the duration of each trial.

Results: Efficacy data were available for 566 subjects who took ramelteon 8?mg (mean age 46.7?years) and 556 subjects who took placebo (mean age 47.8?years). Mean LPS at baseline was 66.6?min for the placebo group and 66.9?min for the ramelteon group. At nights 1 and 2, mean LPS for the ramelteon 8?mg group (30.2?min) was significantly less than the mean LPS for the placebo group (43.3?min). The least squares mean difference from placebo was –13.1?min (p?<?0.001). Headache (8.9% ramelteon 8?mg, 8.8% placebo) and somnolence (3.5% ramelteon 8?mg, 0.7% placebo) were the most common adverse events.

Conclusions: Ramelteon 8?mg, on average, reduced LPS by approximately 13?min more than placebo on nights 1 and 2 of treatment in adults with chronic insomnia. Ramelteon was well tolerated with a low incidence of adverse events. This mean reduction in LPS versus placebo is similar to what has been reported for other classes of insomnia medications. However, these results reflect nights 1 and 2 of treatment and may not be representative of longer treatments.     

α-Flupenthixol increases slow-wave sleep in rats: Effect of dopamine receptor blockade

α-Flupenthixol (0.2 mg/kg, i.p.). a dopamine receptor blocker, significantly increased slow-wave sleep and decreased wakefulness in rats when administered either at the onset of a 12-hr light period or a 12-hr dark period. The same dose of the drug strongly antagonized the dopamine-mediated stereotypy produced by an injection of apomorphine (1 mg/kg, i.p.) for at least 9-hr, indicating that dopamine receptors were blocked throughout most of the recording session. The results suggest a relationship between the blockade of dopamine receptors and the increase of slow-wave sleep time.     

Emerging pharmacotherapeutic agents for insomnia: a hypnotic panacea?

The burden of insomnia has had a significant effect not only on the socioeconomic matrix, but also the medical terrain, as signified by the increased morbidity and mortality of its associated psychiatric and organic sequelae. To this end, a plethora of pharmacotherapeutic agents have been recently introduced that address the vital need to combat insomnia and prevent the perpetuation in its chronic form. The previously and currently dispensed barbiturates and benzodiazepines, respectively, have paved the way for newer agents that are purported to be just as effective, or even more so, with a favourable profile in all domains of sleep. In assessing both published clinical studies and unpublished reports conducted on these emerging agents, this article profiles the most contemporary, therapeutic options in lieu of older hypnotics, over-the-counter medications and supplements. Furthermore, this paper aims to indicate both the future course of hypnotics and the developments currently in progress.     

Honokiol promotes non-rapid eye movement sleep via the benzodiazepine site of the GABA(A) receptor in mice

BACKGROUND AND PURPOSE

Decoctions of the Chinese herb houpu contain honokiol and are used to treat a variety of mental disorders, including depression. Depression commonly presents alongside sleep disorders and sleep disturbances, which appear to be a major risk factor for depression. Here, we have evaluated the somnogenic effect of honokiol and the mechanisms involved.

EXPERIMENTAL APPROACH

Honokiol was administered i.p. at 20:00 h in mice. Flumazenil, an antagonist at the benzodiazepine site of the GABA_A receptor, was administered i.p. 15 min before honokiol. The effects of honokiol were measured by EEG and electromyogram (EMG), c-Fos expression and in vitro electrophysiology.

KEY RESULTS

Honokiol (10 and 20 mg·kg⁻¹) significantly shortened the sleep latency to non-rapid eye movement (non-REM, NREM) sleep and increased the amount of NREM sleep. Honokiol increased the number of state transitions from wakefulness to NREM sleep and, subsequently, from NREM sleep to wakefulness. However, honokiol had no effect on either the amount of REM sleep or EEG power density of both NREM and REM sleep. Honokiol increased c-Fos expression in ventrolateral preoptic area (VLPO) neurons, as examined by immunostaining, and excited sleep-promoting neurons in the VLPO by whole-cell patch clamping in the brain slice. Pretreatment with flumazenil abolished the somnogenic effects and activation of the VLPO neurons by honokiol.

CONCLUSION AND IMPLICATIONS

Honokiol promoted NREM sleep by modulating the benzodiazepine site of the GABA_A receptor, suggesting potential applications in the treatment of insomnia, especially for patients who experience difficulty in falling and staying asleep.     

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。     

Pharmacotherapy of insomnia

Introduction: Insomnia is one of the most prevalent sleep disorders in developed countries, being surpassed only by chronic sleep deprivation. Patients with insomnia tend to have an altered quality of life, impaired daytime functioning and an increased risk of work accidents and motor vehicle crashes. Insomnia is commonly associated with chronic medical conditions, metabolic illnesses and mental disorders (such as depression and anxiety), with which there is a dual, reciprocal relationship.

Areas covered: This paper focuses on current pharmacotherapy options for the treatment of insomnia, particularly benzodiazepine receptor agonists, which nowadays represent the mainstay of hypnotic therapy. The melatonin receptor antagonist, ramelteon, is reviewed (an alternative for some patients with only sleep-onset difficulty), as are sedating antidepressants, which are commonly used ‘off-label’ to treat insomnia, despite limited efficacy data and potential significant safety concerns. Orexin (OX) antagonists are also discussed, especially those that block OX2 or both OX1 and OX2 receptors, as these are the most promising new agents for the treatment of insomnia, with encouraging results in preliminary clinical trials.

Expert opinion: Research to evaluate and formulate treatments for insomnia is often complicated by the fact that insomnia is usually of multifactorial etiology. Understanding the molecular and receptor mechanisms involved in promoting sleep in varied disorders could provide future approaches in new drug development. In the long term, more randomized controlled trials are needed to assess both short-term and long-term effects of these medications and their efficacy in comorbid diseases that affect sleep quality or quantity.     

Package inserts for antihypertensive drugs: use by the patients and impact on adverse drug reactions

Summary The effect of a single dose (30 mg) of Org 3770 (metirzapine) on human sleep was assessed in a double blind, placebo controlled, cross over study in 6 young, healthy male volunteers. The sleep stage classification was based on visual scoring of 24 h electroencephalographic recordings according to the criteria of Rechtschaffen and Kales.Org 3770 30 mg p.o. given 2 h before bedtime had a sleep promoting action in all subjects, resulting in a shortened time to the onset of sleep. Bedtime waking and dozing (Stage 1) were reduced in favour of deep, slow wave sleep (Stages 3 and 4). Org 3770 increased the latency of REM sleep with respect to Stage 2 sleep in all subjects. It also caused a minor reduction in waking periods during REM sleep and a lower frequency of awakenings after periods of movement. No effect of Org 3770 was observed in reaction and vigilance tests on the post treatment day.The observed effects of Org 3770 on normal human sleep suggest that it might ameliorate the sleep disturbances encountered in endogenous depression, which are characterized by a reduction in slow wave sleep, an increase in nighttime awakenings and shortening of REM sleep latency.