本文選題：抑郁癥 + 海馬�。� 參考：《華北理工大學》2015年碩士論文

【摘要】：目的利用慢性不可預見性溫和應激(chronic unpredicted mild stress,CUMS)建立大鼠抑郁癥模型,通過觀察微管相關蛋白p MAP-2和p-Tau在抑郁癥大鼠海馬的蛋白表達變化情況,進而探討其與抑郁癥的關系和意義。方法健康雄性SPF級SD大鼠80只,(體質(zhì)量180~220g)隨機的分成對照組與模型組(n=40),模型組給予慢性不可預見性溫和應激(CUMS)建立模型。應激包括以下11種:潮濕墊料中24 h、進行行為束縛24 h、鼠籠進行45°傾斜24h、4℃的冰水游泳進行5 min、禁水24 h、禁食24 h、42℃熱水游泳進行5 mi n、給予雙耳電擊5 s兩次(0.8 m A)、夾尾1 min、進行鼠籠晃動15 min、持續(xù)光照明24 h。以上11種應激隨機的進行分配,每天給予大鼠1種應激刺激,同種應激刺激不連續(xù)出現(xiàn),共計兩輪。對照組大鼠除每天抓取一次外,不做特殊處理。在應激刺激結束后分別于1、7、14和28d處死大鼠,對照組大鼠處死時間和模型組相同。分別通過糖水偏好實驗、曠場實驗以及Morris水迷宮實驗對對照組和模型組大鼠進行行為學的檢測;尼氏染色觀察大鼠的海馬神經(jīng)元形態(tài)學;透射電鏡觀察大鼠海馬神經(jīng)元細胞超微結構,特別是微管結構的變化;采用免疫熒光雙標法觀察大鼠海馬組織中p MAP-2和p-Tau共定位表達;采用Western blotting方法檢測大鼠海馬組織中p MAP-2和p-Tau蛋白表達水平。采用SPSS17.0統(tǒng)計軟件對數(shù)據(jù)進行統(tǒng)計分析,以P0.05表示差異有統(tǒng)計學意義。結果糖水偏好實驗:對照組和模型組大鼠普通水消耗量無明顯變化,模型組大鼠糖水消耗量和糖水偏好百分比分別低于對照組,差異具有統(tǒng)計學的意義。曠場實驗:模型組大鼠行走的總路程、中央格活動的時間、直立的次數(shù)和修飾行為次數(shù)少于對照組(P0.01)。Morris水迷宮實驗:模型組大鼠的平均逃避潛伏期高于對照組(P0.01)。尼氏染色:結果表現(xiàn)為對照組的大鼠海馬細胞密度大,錐體細胞層厚,排列緊密整齊,胞體飽滿,突起顯明。模型組大鼠的海馬細胞密度變小,錐體細胞層變薄,細胞間隙變大,排列分布疏松,細胞核萎縮,突起相對減少。透射電鏡:結果表現(xiàn)為對照組大鼠海馬的神經(jīng)細胞核膜較為完整,染色質(zhì)結構相對正常,超微結構正常。模型組海馬的細胞核相對固縮,核異染色質(zhì)增多,發(fā)現(xiàn)一些小而不規(guī)則凝塊、邊緣聚合,內(nèi)質(zhì)網(wǎng)腫脹,線粒體嵴消失、腫脹、空泡化,還有細胞器減少、結構遭到破壞,以及在胞漿中發(fā)現(xiàn)形成大小不等的空泡。免疫熒光雙標結果:各組大鼠海馬區(qū)p MAP-2、p-Tau檢測結果:p MAP-2為Alexa Fluor 488激發(fā)的綠色熒光,p-Tau為Alexa Fluor 594激發(fā)的紅色熒光,圖像重疊后可見部分神經(jīng)元的胞漿內(nèi)p MAP-2和p-Tau同時表達,表明p MAP-2和p-Tau有部分共定位。Western blotting檢測結果:根據(jù)條帶分析顯示與對照組比較,p MAP-2蛋白和p-T au蛋白分別在CUMS后1d蛋白表達低于對照組(P0.01),7 d后開始升高(P0.01),14 d明顯高于對照組(P0.01),28 d仍略高于對照組(P0.05)。結論1成功建立抑郁癥的動物模型。2抑郁模型大鼠海馬神經(jīng)元發(fā)生變化。3微管蛋白p MAP-2和p-Tau蛋白表達水平在抑郁模型大鼠海馬先降低后增高變化,這種變化可能影響微管的穩(wěn)定性,破壞細胞骨架穩(wěn)態(tài),從而影響海馬神經(jīng)元的結構改變。
[Abstract]:Objective to establish a rat model of depression with chronic unpredictability and mild stress (chronic unpredicted mild stress, CUMS), and to explore the relationship and significance of the protein expression of microtubule related protein P MAP-2 and p-Tau in the hippocampus of depressive rats, and to explore the relationship and significance of the protein expression of P MAP-2 and p-Tau in the hippocampus of the depressive rats. Methods 80 healthy male SPF SD rats, (physique) 180~220g) was randomly divided into the control group and the model group (n=40). The model group was given a model of chronic unpredictability and mild stress (CUMS). The stress included 11 kinds: 24 h in the damp mat, 24 h for behavior, 45 degrees in the squirrel cage, 5 min for swimming at 4 C, 24 h, 24 h, and 5 Mi n in hot water at 42. The double ear electric shock was given 5 s two times (0.8 m A) and tail 1 min, the squirrel cage sloshing 15 min, and 11 kinds of stress above 24 h. in the continuous light illumination were randomly assigned. The rats were given 1 stress stimuli every day. The same stress irritation was discontinuous in two rounds. The control group was taken once a day and did not do special treatment. After the end of the stress stimulation, the rats were not treated with special treatment. The rats were killed in 1,7,14 and 28d respectively. The rats in the control group were killed at the same time as the model group. The behavior of the rats in the control group and the model group were detected by the sugar water preference experiment, the open field experiment and the Morris water maze experiment. The hippocampus neuron morphology was observed by Nissl staining, and the hippocampal neurons were observed by transmission electron microscope. The ultrastructure of the cell, especially the microtubule structure, was used to observe the co localization of P MAP-2 and p-Tau in the hippocampus of rats by immunofluorescence, and the Western blotting method was used to detect the expression of P MAP-2 and p-Tau protein in the hippocampus of rats. The data were statistically analyzed by the SPSS17.0 statistical software, and the difference was expressed by P0.05. Results the sugar water preference experiment: the normal water consumption of the control group and the model group had no obvious change. The sugar water consumption and the sugar water preference percentage in the model group were lower than those of the control group. The difference was statistically significant. The open field experiment: the total distance of walking in the model group, the time of the central grid activity, the erect times and the number of the rats in the model group. The number of modified behavior was less than that of the control group (P0.01).Morris water maze experiment: the average escape latency of the model group was higher than that of the control group (P0.01). Nissl staining: the results showed that the hippocampus cell density of the control group was large, the pyramidal cell layer was thick, the cell body was full and the protuberance was obvious. The density of the hippocampus cells in the model group became smaller. The pyramidal cell layer became thinner, the cell space became larger, the distribution was loose, the nucleus atrophied and the protuberance was relatively reduced. Transmission electron microscope showed that the nuclear membrane of the hippocampus of the control group was more complete, the chromatin structure was relatively normal, and the ultrastructure was normal. The nucleus of the model group was relatively condensed, and the nuclear heterochromatin was increased, found that the nuclear heterochromatin was on the increase, found that the nucleus of the nucleus increased. Small and irregular clot, edge polymerization, endoplasmic reticulum swelling, mitochondrial crista disappearance, swelling, vacuolization, organelle reduction, structure destruction, and the discovery of vacuoles in the cytoplasm. Double immunofluorescence results: P MAP-2, p-Tau test results in hippocampus of rats in each group: P MAP-2 for Alexa Fluor 488 green fluores. Light, p-Tau is the red fluorescence excited by Alexa Fluor 594, and the P MAP-2 and p-Tau in the cytoplasm of the visible part of the neurons are expressed at the same time after the image overlap. It shows that P MAP-2 and p-Tau have a partial CO localization of.Western blotting detection results. In the control group (P0.01), after 7 d (P0.01), 14 d was significantly higher than that of the control group (P0.01), and 28 d was still slightly higher than the control group (P0.05). Conclusion 1 successfully established the animal model of depression, the hippocampus neurons of the depressive model of.2, the.3 microtubule P MAP-2 and the expression of the p-Tau protein expression in the hippocampus of the depressive model rats increased and then increased. This change may affect the stability of microtubules and destroy the homeostasis of cytoskeleton, thus affecting the structural changes of hippocampal neurons.

【學位授予單位】：華北理工大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R749.4

【參考文獻】

相關期刊論文 前5條

1 婁冉;牛森林;;抑郁癥與焦慮癥的臨床辨析[J];甘肅中醫(yī);2009年02期

2 王海濤;呂翠平;趙毓芳;魏靜波;闞泉;高俊玲;;抑郁模型大鼠杏仁核神經(jīng)元凋亡及Bax/Bcl-2的變化[J];神經(jīng)解剖學雜志;2010年04期

3 王海濤;徐愛軍;闞泉;李冉;;應激性抑郁癥大鼠杏仁核微管相關蛋白表達的實驗研究[J];神經(jīng)解剖學雜志;2012年02期

4 高杉;李苒;;抑郁癥動物模型及其評價標準研究進展[J];天津中醫(yī)藥大學學報;2012年01期

5 劉福友,楊石,陳衛(wèi)垠,唐熠;郁樂疏對卒中后抑郁模型大鼠神經(jīng)遞質(zhì)的影響研究[J];實用醫(yī)院臨床雜志;2004年02期

相關博士學位論文 前1條

1 蔡同建;tau蛋白磷酸化在錳誘導的神經(jīng)毒性中的作用[D];第四軍醫(yī)大學;2009年

相關碩士學位論文 前1條

1 朱周靚;抑制自噬相關基因表達導致果蠅的運動障礙[D];中南大學;2011年

，

本文編號：1814622