發(fā)布時(shí)間：2018-03-10 11:31

  本文選題：淀粉樣前體蛋白可溶性α片段　切入點(diǎn)：睡眠-覺(jué)醒周期　出處：《第二軍醫(yī)大學(xué)》2012年博士論文　論文類型：學(xué)位論文

【摘要】：【目的】淀粉樣前體蛋白（APP）經(jīng)淀粉樣代謝途徑產(chǎn)生的可溶性β淀粉樣蛋白（Aβ）不正常的聚集與阿爾茨海默�。ˋD）密切相關(guān)。睡眠剝奪影響認(rèn)知功能，長(zhǎng)期睡眠限制的APP轉(zhuǎn)基因鼠可以出現(xiàn)病理Aβ沉積，提示睡眠缺失有可能通過(guò)影響APP代謝途徑參與AD的致病過(guò)程。生理?xiàng)l件下，APP主要經(jīng)非淀粉樣代謝途徑產(chǎn)生可溶性α片段（sAPPα）并發(fā)揮生理保護(hù)作用。在近年多領(lǐng)域針對(duì)APP淀粉樣代謝途徑臨床干預(yù)研究未獲得預(yù)期效果的背景下，重新審視和研究APP非淀粉樣代謝途徑產(chǎn)物sAPPα是否存在與睡眠相關(guān)的變化以及在睡眠剝奪動(dòng)物模型中是否發(fā)揮認(rèn)知保護(hù)機(jī)制具有一定現(xiàn)實(shí)意義。觀察sAPPα在基于睡眠改善的臨床干預(yù)研究中的變化及相關(guān)認(rèn)知能力改變，有可能為認(rèn)知障礙疾病探尋新的干預(yù)或治療靶點(diǎn)。 【方法】建立正常睡眠-覺(jué)醒模型和睡眠剝奪模型，觀察實(shí)驗(yàn)動(dòng)物是否存在與睡眠相關(guān)的sAPPα變化，以及sAPPα是否能夠?qū)顾邉儕Z引起的認(rèn)知功能損害；進(jìn)而觀察慢性失眠患者和AD患者藥物干預(yù)睡眠缺失前后的sAPPα變化以及相應(yīng)的認(rèn)知功能評(píng)分改變。所有涉及本課題的嚙齒類動(dòng)物在模擬外界光暗變化（12h:12h）的條件下適應(yīng)性飼養(yǎng)2周，建立正常的睡眠-覺(jué)醒周期。首先，利用寰枕膜穿刺技術(shù)在24h內(nèi)按時(shí)間節(jié)點(diǎn)（間隔4h）獲取正常睡眠-覺(jué)醒周期Sprague-Dawley（SD）大鼠腦脊液（CSF）樣本，利用免疫印跡技術(shù)（western-blot）檢測(cè)腦脊液sAPPα的水平，判斷其是否存在晝夜節(jié)律性變化。其次，給予上述實(shí)驗(yàn)動(dòng)物12h睡眠剝奪以及12h恢復(fù)性睡眠，獲取此兩節(jié)點(diǎn)CSF樣本，同樣利用免疫印跡技術(shù)觀察急性睡眠剝奪與睡眠恢復(fù)對(duì)sAPPα的影響。此外，利用同上的方法觀察亞急性部分性睡眠剝奪C57BL6小鼠腦內(nèi)sAPPα的影響。第三，觀察SD大鼠急性睡眠剝奪對(duì)認(rèn)知水平（Morris水迷宮空間記憶）的影響，以及腦內(nèi)皮質(zhì)和海馬sAPPα的變化，進(jìn)而觀察腦內(nèi)注射重組sAPPα是否可以對(duì)抗睡眠剝奪對(duì)Morris水迷宮空間記憶的影響。第四，通過(guò)癥狀詢問(wèn)、查體、量表評(píng)估等手段完成AD患者受試對(duì)象和慢性失眠患者受試對(duì)象的納入工作，匹配健康對(duì)照組。針對(duì)各組進(jìn)行認(rèn)知量表評(píng)定、夜間多導(dǎo)睡眠監(jiān)測(cè)（PSG），并在知情同意后行腰椎穿刺獲取腦脊液標(biāo)本。分析各組認(rèn)知評(píng)分、PSG監(jiān)測(cè)數(shù)據(jù)以及腦脊液sAPPα之間的差異，給予右佐匹克�。�3mg/d，2周）治療AD患者和慢性失眠患者的失眠癥狀，觀察治療前后失眠緩解的情況以及相應(yīng)的認(rèn)知評(píng)分和腦脊液sAPPα變化。 【結(jié)果】本研究的結(jié)果顯示（1）SD大鼠24h內(nèi)自然睡眠-覺(jué)醒周期中的6個(gè)時(shí)間節(jié)點(diǎn)腦脊液sAPPα水平發(fā)生具有統(tǒng)計(jì)學(xué)意義的波動(dòng)性變化（p=0.013）。其中，sAPPα水平于睡眠始發(fā)約4h后達(dá)到高峰，而后逐漸下降，覺(jué)醒時(shí)維持相對(duì)穩(wěn)定的水平。（2）SD大鼠在急性完全睡眠剝奪12h后腦脊液中sAPPα水平下降，在恢復(fù)睡眠12h后上升（p=0.024），與環(huán)境對(duì)照之間具有顯著性區(qū)別（p=0.011）。（3）與正常對(duì)照組和環(huán)境對(duì)照組相比，C57BL6小鼠在經(jīng)歷亞急性部分性睡眠剝奪2周后腦組織中無(wú)論是皮質(zhì)還是海馬sAPPα水平均有統(tǒng)計(jì)學(xué)意義的下降（p0.05），但皮質(zhì)與海馬之間sAPPα的濃度無(wú)統(tǒng)計(jì)學(xué)意義上的差異（p0.05）。（4）睡眠剝奪可以影響SD大鼠Morris水迷宮空間記憶（目標(biāo)象限游泳時(shí)間百分比）能力（p=0.000），腦室注射重組sAPPα可以改善空間記憶的損害（p=0.037），這種改善在2次注射后更加顯著（p=0.005）。（5）與健康對(duì)照相比，AD患者和慢性失眠患者均有睡眠效率減低（p=0.000），睡眠期間覺(jué)醒增加（p=0.000）以及REM睡眠減少（p=0.000）；AD患者覺(jué)醒時(shí)間較慢性失眠患者增加（p=0.018），，而慢性失眠患者睡眠潛伏期明顯延長(zhǎng)（p=0.000）。（6）AD患者腦脊液sAPPα水平較慢性失眠患者和健康對(duì)照下降（p=0.000，p=0.000），后兩者之間無(wú)顯著差異（p0.05）。（7）AD患者腦脊液sAPPα水平與認(rèn)知量表評(píng)分和睡眠效率之間具有顯著的正相關(guān)（spearman’s rho0.676, p=0.016;0.935, p=0.000）。（8）右佐匹克隆干預(yù)治療失眠可以明顯改善PSG睡眠參數(shù)（p0.05）。（9）右佐匹克隆干預(yù)可以改善慢性失眠患者認(rèn)知量表評(píng)分（p=0.047），但不改變腦脊液sAPPα水平（p=0.142）；干預(yù)治療可以增加AD患者腦脊液sAPPα水平（p=0.004），但AD患者認(rèn)知量表評(píng)分沒(méi)有顯著改善（p=0.514）。 【結(jié)論】首先，sAPPα在正常睡眠-覺(jué)醒狀態(tài)下呈節(jié)律性變化，在睡眠期達(dá)到高峰。無(wú)論是急性完全睡眠剝奪還是亞急性部分性睡眠剝奪均可導(dǎo)致腦內(nèi)sAPPα水平的下降，急性睡眠剝奪導(dǎo)致的下降在恢復(fù)性睡眠之后可以迅速恢復(fù)，提示睡眠與腦內(nèi)sAPPα水平具有相關(guān)性。其次，腦內(nèi)注射重組sAPPα可以對(duì)抗急性完全睡眠剝奪動(dòng)物模型中空間記憶的損害。第三，AD患者腦脊液中sAPPα水平下降，與認(rèn)知評(píng)分及睡眠時(shí)間呈正相關(guān)，同樣存在失眠癥狀的慢性失眠患者sAPPα水平正常。藥物干預(yù)改善失眠癥狀后，AD患者腦脊液sAPPα水平增加，但認(rèn)知評(píng)分無(wú)顯著恢復(fù)，而針對(duì)失眠的干預(yù)治療可以改善慢性失眠患者的認(rèn)知評(píng)分。說(shuō)明人體中腦內(nèi)sAPPα水平與睡眠存在相關(guān)性，改善睡眠可以增加腦內(nèi)sAPPα水平，但是對(duì)于已經(jīng)發(fā)展成阿爾茨海默病的患者，睡眠的改善不能逆轉(zhuǎn)認(rèn)知功能缺損。
[Abstract]:[Objective] the amyloid precursor protein (APP) soluble amyloid beta protein produced by amyloid metabolic pathways (A beta) aggregation and Blzheimer's disease is not normal (AD) are closely related. The effects of sleep deprivation on cognitive function, long-term sleep restriction of APP transgenic mice can appear pathological A deposition, suggesting that lack of sleep may the pathogenic effects of APP metabolic pathway in AD. Under physiological conditions, APP mainly by non amyloid metabolic pathways to produce soluble fragment of alpha (sAPP alpha) and its physiological protective effect. In recent years many fields in APP amyloid metabolic pathways of clinical intervention did not achieve the expected effect of the background, review and study of APP non amyloid metabolism product of sAPP alpha if there are changes related to sleep and sleep deprivation in animal models of cognitive play protection mechanism has a certain practical significance. Based on the observation of sAPP alpha Changes in the clinical intervention study of sleep improvement and related cognitive changes may be possible to explore new interventions or targets for cognitive impairment.
[Methods] to establish normal sleep wake model and sleep deprivation model, observe the experimental animal existence of sAPP alpha changes and sleep related, and whether the sAPP alpha against cognitive impairment caused by sleep deprivation; and observation of patients with chronic insomnia and drug intervention in patients with AD sAPP alpha changes before and after sleep deprivation and the corresponding cognitive function score change. All refer to this paper in rodent animal simulation of external light dark changes (12h:12h) under the condition of adaptive feeding for 2 weeks, the establishment of normal sleep wake cycle. First of all, the atlanto occipital membrane puncture technique according to the time node within 24h (4h interval) to get normal sleep wake cycle Sprague-Dawley (SD) rat cerebrospinal fluid (CSF) samples by Western blot (Western-blot) levels in cerebrospinal fluid of sAPP alpha, to determine whether it has circadian change. Secondly, given the above experiment Animal 12h sleep deprivation and 12h recovery sleep, get the two node CSF samples, the same observation of acute sleep deprivation and recovery effects on sAPP alpha sleep by Western blot. In addition, the observation of subacute part of the impact of sleep deprivation in C57BL6 mice brain using sAPP alpha ibid method. Third, observation of acute sleep SD after deprivation on cognitive level (Morris water maze memory space) effects, and changes in the brain cortex and hippocampus of sAPP alpha, and observe whether intracerebral injection of recombinant alpha sAPP can counteract the effects of sleep deprivation on Morris water maze spatial memory. Fourth, inquiry, examination by symptoms scale assessment method AD subjects and patients with chronic insomnia subjects included in the work, the healthy control group. The cognitive assessment scales for each group, nocturnal polysomnography (PSG), and informed consent was waist Lumbar puncture CSF samples obtained were analyzed. The differences between cognitive scores, PSG monitoring data and cerebrospinal fluid of sAPP alpha, given dexzopiclon (2 weeks 3mg/d) treatment of patients with AD and patients with chronic insomnia insomnia symptoms were observed before and after treatment of insomnia relief and the corresponding cognitive score and cerebrospinal fluid sAPP alpha changes.
[result] the results of this study show that (1) natural sleep SD rats 24h awakening cycle in 6 time nodes of cerebrospinal fluid sAPP levels fluctuate change was statistically significant (p=0.013). The levels of sAPP in sleep onset after about 4H reached the peak, then decreased gradually, to maintain a relatively stable the level of awakening. (2) sAPP in cerebrospinal fluid of acute 12h after total sleep deprivation in the alpha level of SD rats decreased, increased in sleep recovery after 12h (p=0.024), with a significant difference between the control and the environment (p=0.011). (3) compared with the normal control group and control group, C57BL6 mice after subacute partial sleep deprivation for 2 weeks in both cerebral cortex and hippocampus decreased sAPP levels were statistically significance (P0.05), but no statistically significant difference between the concentrations of cortex and hippocampus sAPP alpha (P0.05). (4) sleep deprivation can influence S D rats in Morris water maze spatial memory (the percentage of swimming time in the target quadrant) capacity (p=0.000), intraventricular injection of recombinant alpha sAPP can improve the spatial memory impairment (p=0.037), this improvement is more significant in 2 after injection (p=0.005). (5) compared with healthy controls, reduce AD patients and patients with chronic insomnia were sleep efficiency (p=0.000), increased awakening during sleep (p=0.000) and REM (p=0.000); sleep decreased in patients with AD increased the awakening time of patients with chronic insomnia (p=0.018), and patients with chronic insomnia sleep latency was significantly prolonged (p=0.000). (6) AD in cerebrospinal fluid of patients with sAPP levels than patients and healthy controls (decrease of chronic insomnia p=0.000, p=0.000), no significant difference between (P0.05). (7) sAPP alpha level and cerebrospinal fluid in patients with AD cognitive scale score and sleep efficiency has a significant positive correlation (Spearman 's rho0.676, p=0.016; 0.935, p=0.000 (8). ) dexzopiclone intervention in the treatment of insomnia can significantly improve the sleep parameters of PSG (P0.05). (9) dexzopiclone intervention can improve the cognition of patients with Chronic Insomnia Scale score (p=0.047), but does not change the CSF levels of sAPP (p=0.142); intervention therapy can increase AD cerebrospinal fluid levels of sAPP alpha (p=0.004). But the cognition of the patients with AD score did not improve significantly (p=0.514).
[Conclusion] first, sAPP alpha showed circadian changes in normal sleep wake state, reached the peak in the sleep period. Both acute or subacute complete sleep deprivation of sleep deprivation can lead to decrease sAPP levels in the brain, acute sleep deprivation led to decreased can be rapidly recovered after re sleep after recovery, has the correlation suggested that sAPP alpha level sleep and the brain. Secondly, intracerebral injection of recombinant sAPP alpha can memory against acute total sleep deprivation in the animal model of damage. Third, the levels of sAPP alpha in cerebrospinal fluid of AD patients was positively correlated with cognitive decline, score and sleep time, there are also symptoms of insomnia insomnia patients with chronic sAPP alpha level normal. Drug intervention to improve the symptoms of insomnia, AD in cerebrospinal fluid of patients with sAPP levels increased, but cognitive score no significant recovery, but for insomnia intervention can improve the treatment of chronic insomnia patients The score of cognition indicates that there is a correlation between the level of sAPP alpha in the human brain and sleep. The improvement of sleep can increase the level of sAPP alpha in the brain. But for those who have developed Alzheimer's disease, the improvement of sleep can not reverse the cognitive impairment.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R749.16

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 馬哲;王平;游秋云;;睡眠及睡眠剝奪與學(xué)習(xí)記憶的相關(guān)性探討[J];中華中醫(yī)藥雜志;2014年04期

相關(guān)碩士學(xué)位論文 前1條

1 馬哲;《黃帝內(nèi)經(jīng)》神與睡眠關(guān)系的理論探討及益元安神方作用的研究[D];湖北中醫(yī)藥大學(xué);2014年

本文編號(hào)：1593158