本文選題：細胞型朊蛋白 + 睡眠剝奪��； 參考：《第二軍醫(yī)大學》2017年碩士論文

【摘要】：【目的】隨著生活節(jié)奏和工作壓力的增加,睡眠障礙的現(xiàn)象越來越普遍,以往研究表明睡眠障礙是阿爾茨海默病(Alzheimer’s disease,AD)的臨床癥狀之一。目前的研究發(fā)現(xiàn)睡眠障礙不僅是AD的癥狀,還有可能是AD發(fā)病的危險因素。細胞型朊蛋白(Cellular prion protein,PrP~c)是一種高度保守的在中樞神經(jīng)系統(tǒng)高表達的膜蛋白,其生理功能目前尚不清楚,但有研究證實PrPC在睡眠調(diào)節(jié)和記憶形成與鞏固過程發(fā)揮重要作用。有研究顯示小鼠視交叉上核和頂葉皮層PrPC mRNA具有晝夜節(jié)律變化,提示PrP~c可能存在晝夜節(jié)律性;有研究發(fā)現(xiàn)PrP~c可抑制關鍵酶β-分泌酶(β-secretase,BACE1)活性,調(diào)節(jié)APP的分解,從而影響Aβ的產(chǎn)生。另外,研究已經(jīng)證實睡眠剝奪在導致認知功能損害的同時伴隨Aβ表達上調(diào),提示睡眠剝奪誘導認知功能損害可能與PrP~c對Aβ的影響有關,PrP~c可能具有潛在神經(jīng)保護作用。探討PrP~c在睡眠剝奪所致認知損害中發(fā)揮的作用,有可能為認知障礙疾病找到潛在治療靶點。【方法】選取C57B L/6小鼠,按體重大小隨機分成6組,正常飼養(yǎng)2周以建立正常睡眠-覺醒周期,分別于24h內(nèi)6個不同時間點獲取取海馬和皮層組織樣本,采用ELISA方法檢測PrP~c與Aβ的表達水平變化,以判斷是否存在晝夜節(jié)律變化。其次,選取C57B L/6小鼠,隨機分成CC組、EC組、SD組,采用改良水平臺法進行72h睡眠剝奪后獲取海馬組織,采用Western Blot檢測PrP~c和ELISA方法檢測Aβ表達變化以觀察睡眠剝奪后小鼠海馬PrP~c與Aβ的表達情況。第三,采用腦立體定位注射技術在小鼠海馬注射PrP~c過表達腺相關病毒,觀察PrP~c過表達是否能改善睡眠剝奪引起的小鼠空間記憶損害。【結果】本研究結果顯示(1)C57B L/6小鼠腦內(nèi)Aβ在24小時正常睡眠-覺醒周期中6個不同時間點出現(xiàn)有統(tǒng)計學意義的波動性變化(P0.05);單余弦分析結果顯示PrPC在皮層部位存在晝夜節(jié)律性(F=11.22,P0.05)。Aβ在海馬部位具有晝夜節(jié)律性(F=26.72,P0.05)。(2)睡眠剝奪后海馬PrPC的表達在SD組(0.22±0.05)較CC組(0.64±0.16)和EC組(0.58±0.09)明顯下調(diào)(F=4.366,P0.05);Aβ的表達在SD組(13.03±0.71)較CC組(8.22±0.8)和EC組(8.6±0.57)明顯上調(diào)(F=14.511,P0.05)。(3)海馬內(nèi)注射腺相關病毒使PrP~c過表達可以改善睡眠剝奪誘導的海馬空間記憶損害,過表達組目標象限時間百分比顯著高于對照組(F=6.196,P0.05)�！窘Y論】首次發(fā)現(xiàn),小鼠皮層PrPC的表達在正常睡眠-覺醒周期下表現(xiàn)為晝夜節(jié)律變化,在覺醒期達到高峰,急性睡眠剝奪可導致海馬內(nèi)PrPC表達下降,提示小鼠腦內(nèi)PrPC表達水平與睡眠相關。其次,海馬內(nèi)PrPC過表達可以改善睡眠剝奪后空間記憶損害。提示PrPC具有潛在神經(jīng)保護作用。
[Abstract]:[objective] with the increase of life rhythm and work stress, sleep disorder is becoming more and more common. Previous studies have shown that sleep disorder is one of the clinical symptoms of Alzheimer's disease (AD). Current studies have found that sleep disorder is not only a symptom of AD, but also a risk factor for AD. Cellular prion protein (prion) is a highly conserved membrane protein expressed in the central nervous system (CNS), and its physiological function is not clear, but it has been confirmed that it plays an important role in the process of sleep regulation and memory formation and consolidation. Some studies have shown that PrPC mRNA in suprachiasmatic nucleus and parietal cortex has circadian rhythm, suggesting that PrPnc may have circadian rhythm, and it has been found that PrPcc can inhibit the activity of 尾 -secretase (尾 -secretase) and regulate the decomposition of app, thus affecting the production of A 尾. In addition, it has been confirmed that sleep deprivation may lead to cognitive impairment accompanied by up-regulation of A 尾 expression, suggesting that sleep deprivation induced cognitive impairment may be related to the effect of PrPnc on A 尾 and may have a potential neuroprotective effect. To explore the role of PrPtroc in cognitive impairment induced by sleep deprivation, it is possible to find a potential therapeutic target for cognitive impairment. [methods] C57B / L / 6 mice were randomly divided into 6 groups according to their body weight. The normal sleep-wake cycle was established by normal feeding for 2 weeks. The hippocampal and cortical tissues were collected at 6 different time points within 24 hours. The expression levels of PrPnc and A 尾 were detected by Elisa to determine whether there were circadian rhythms. Secondly, C57B / L / 6 mice were randomly divided into CC group (EC group) and SD group. Hippocampal tissue was obtained after 72 h sleep deprivation by modified horizontal table method. The expression of A 尾 was detected by Western blot and Elisa in order to observe the expression of PrPnc and A 尾 in hippocampus of mice after sleep deprivation. Thirdly, using stereotactic injection technique, we injected PrPnc overexpression adeno-associated virus into the hippocampus of mice. [results] the present study showed that (1) A 尾 in the brain of C57B / L / 6 mice appeared at 6 different time points in the normal sleep / arousal cycle of 24 hours. [results] the results showed that A 尾 in the brain of C57B / L / 6 mice was regulated at 6 different time points during the normal sleep / wake-up cycle of 24 hours. The results of single cosine analysis showed that there was circadian rhythm in cortical area (F _ (11.22) P _ (0.05) .A 尾 in hippocampus (F _ (26.72) P _ (0.05). (_ 2). The expression of PrPC in hippocampus after sleep deprivation was (0.22 鹵0. 05) in SD group compared with (0.64 鹵0.16) in CC group and (0.58 鹵0.09) in EC group. In SD group (13.03 鹵0.71), compared with CC group (8.22 鹵0.8) and EC group (8.6 鹵0.57), the down-regulated expression of). (尾 was significantly up-regulated (F _ (14. 511) P 0.05 /). (_ 3). The overexpression of PrPc in hippocampus induced by sleep deprivation could improve the spatial memory impairment of hippocampus induced by sleep deprivation, and the expression of PrPc 尾 was significantly up-regulated in SD group (P < 0.05), compared with that in CC group (8.22 鹵0.8) and EC group (8.6 鹵0.57). The percentage of target quadrant time in the overexpression group was significantly higher than that in the control group (P 0.05). [conclusion] it was found for the first time that the expression of PrPC in the cortex of mice showed circadian rhythm changes during normal sleep-wake cycle, and reached its peak at the awakening stage. Acute sleep deprivation resulted in a decrease in the expression of PrPC in the hippocampus, suggesting that the expression level of PrPC in the brain of mice was related to sleep. Secondly, overexpression of PrPC in hippocampus can improve spatial memory impairment after sleep deprivation. The results suggest that PrPC has potential neuroprotective effect.
【學位授予單位】：第二軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R740

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中國期刊全文數(shù)據(jù)庫 前1條

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本文編號：2096452