本文選題：邊緣下區(qū) + NR2B��； 參考：《第三軍醫(yī)大學(xué)》2010年碩士論文

【摘要】： 背景:近年來(lái),隨著各種突發(fā)災(zāi)難性事件(地震、洪水、恐怖襲擊、車禍等)的發(fā)生,創(chuàng)傷后應(yīng)激障礙(post-traumatic stress disorder,PTSD)成了關(guān)注的重點(diǎn)。所謂PTSD是親身經(jīng)歷或者目睹異乎尋常的威脅性、災(zāi)難性事件后發(fā)生的一系列生理、心理應(yīng)激所表現(xiàn)出來(lái)的臨床綜合癥。創(chuàng)傷記憶一旦形成,就很難消退,給患者帶來(lái)很大痛苦。目前臨床上治療PTSD常用的非藥物方法為暴露療法(基于消退原理的行為治療方法),但約有40%的病人療效難以維持,表現(xiàn)為已消退的恐懼記憶很快再現(xiàn)。這提示消退保持障礙可能是PTSD發(fā)生和治療困難的關(guān)鍵。因此,研究恐懼消退保持的神經(jīng)生物學(xué)機(jī)制具有重要意義。目前,相對(duì)于條件性恐懼的獲得、鞏固與表達(dá)過(guò)程,對(duì)恐懼消退機(jī)制的研究尚很不夠,許多問(wèn)題有待闡明。 恐懼消退是指當(dāng)條件性刺激(conditioned stimulus,CS)重復(fù)呈現(xiàn)而缺乏非條件刺激(unconditioned stimulus,US,如足底電擊)配對(duì)呈現(xiàn),所致的條件性恐懼反應(yīng)的逐漸減弱或消失。它不是抹掉了先前形成的CS-US聯(lián)結(jié)記憶,而是形成了CS-no US的新記憶(消退記憶)。大量研究表明內(nèi)側(cè)前額葉(medial prefrontal cortex,mPFC)邊緣下區(qū)(infralimbic cortex,IL)參與了條件化恐懼的消退過(guò)程,在消退保持中起重要作用,是消退記憶長(zhǎng)期保持的場(chǎng)所。但是IL區(qū)在消退記憶長(zhǎng)時(shí)保持中的作用機(jī)制如何?哪些物質(zhì)參與了消退過(guò)程尚不清楚。 我們?cè)谝酝芯康幕A(chǔ)上,選用條件性恐懼模型和恐懼消退模型,綜合行為學(xué)、形態(tài)學(xué)和Western-Blot方法對(duì)大鼠IL區(qū)在恐懼消退保持中的作用及機(jī)制進(jìn)行探索,以期找到條件性恐懼消退的最佳“時(shí)間窗”和關(guān)鍵分子,探討IL區(qū)在聲音條件性恐懼消退保持中的作用機(jī)制,為提高PTSD治療效果提供依據(jù)。 目的:探討給予條件性恐懼消退訓(xùn)練后,大鼠邊緣下區(qū)NR2B、BDNF以及caspase-3在消退保持中的動(dòng)態(tài)變化。 方法:75只SD大鼠隨機(jī)分成空白對(duì)照組(Cont組,無(wú)任何處理),消退對(duì)照組(N-EXE組,無(wú)消退訓(xùn)練)和24h消退組(EXE組)。采用聲音-足底電擊配對(duì)刺激建立條件性恐懼模型,并于恐懼建立24h后進(jìn)行消退訓(xùn)練,比較消退訓(xùn)練后第1、3、7天大鼠的僵立行為變化以及IL區(qū)NR2B、BDNF以及caspase-3的變化。 結(jié)果: 1.各組大鼠消退保持成績(jī)的變化: 消退訓(xùn)練后,消退組大鼠消退保持成績(jī)逐漸增加(恐懼逐漸減少)。消退訓(xùn)練后第3天和7天,24h消退組消退保持成績(jī)好于消退對(duì)照組。(p0.01)。 2.各組大鼠IL區(qū)NR2B、BDNF免疫反應(yīng)陽(yáng)性物質(zhì)的變化: (1)消退對(duì)照組和24h消退組大鼠IL區(qū)NR2B和BDNF含量均逐漸增加,第1、3.天增加迅速,第7天最高,但均低于空白對(duì)照組,差異顯著(p0.01); (2)消退對(duì)照組和24h消退組相比較:第1天兩組無(wú)明顯差異;第3、7天兩組差異顯著,且24h消退組顯著高于消退對(duì)照組(p0.05)。 3.各組大鼠mPFC NR2B以及BDNF蛋白含量的變化: 消退對(duì)照組和24h消退組大鼠在消退后第1、3、7天IL區(qū)NR2B、BDNF蛋白含量逐漸增加,第1、3天增加迅速,消退對(duì)照組第7天基本恢復(fù)正常。在各時(shí)間點(diǎn),消退對(duì)照組NR2B和BDNF的蛋白含量顯著低于24h消退組和空白對(duì)照組。 4.各組大鼠IL區(qū)caspase-3免疫反應(yīng)陽(yáng)性物質(zhì)的變化: 與空白對(duì)照組比較,(1)消退對(duì)照組和24h消退組大鼠IL區(qū)caspase-3免疫反應(yīng)陽(yáng)性物質(zhì)量明顯高于空白對(duì)照組;消退訓(xùn)練后第7天,24h消退組caspase-3免疫反應(yīng)陽(yáng)性物質(zhì)量接近空白對(duì)照組水平;(2)消退訓(xùn)練后隨著時(shí)間的增加,消退對(duì)照組和24h消退組caspase-3含量逐漸減少,第7天24h消退組caspase-3量基本恢復(fù)正常;(3)在各時(shí)間點(diǎn),24h消退組caspase-3量顯著低于消退對(duì)照組(p0.05)。 結(jié)論: 1.行為學(xué)的結(jié)果顯示:恐懼消退訓(xùn)練后一周內(nèi),消退對(duì)照組和24h消退組大鼠的消退保持成績(jī)均逐漸提高(條件性恐懼逐漸改善),24h消退組好于消退對(duì)照組。提示:消退訓(xùn)練有利于恐懼記憶的消退。因此,對(duì)PTSD患者要給予及時(shí)積極的暴露治療; 2.免疫組化和Western Blot結(jié)果顯示: IL區(qū)NR2B和BDNF的免疫陽(yáng)性細(xì)胞數(shù)以及蛋白水平隨著消退成績(jī)的改善而逐漸增加,在消退訓(xùn)練后3-7天達(dá)最高。提示:消退訓(xùn)練后3-7天可能是恐懼消退保持的關(guān)鍵時(shí)段,IL區(qū)NR2B和BDNF在恐懼消退保持中起重要作用。 3.消退訓(xùn)練后,隨著IL區(qū)caspase-3含量逐漸下降,且24h消退組成績(jī)好于消退對(duì)照組。提示:caspase-3可能參與了恐懼消退記憶保持過(guò)程中IL區(qū)的可塑性變化,但其中的機(jī)制還需進(jìn)一步研究。
[Abstract]:Background: in recent years, post-traumatic stress disorder (PTSD) has become a focus with the occurrence of a variety of catastrophic events (earthquakes, floods, terrorist attacks, and car accidents). The so-called PTSD is a series of physiological, psychological stresses that occur after the experience of personal experience or the unusual coerced, disastrous events. The manifestation of clinical syndrome. Once the traumatic memory is formed, it is difficult to recede and bring great pain to the patient. At present, the clinical treatment of PTSD is a non drug method for exposure therapy (based on the behavior therapy based on the principle of regression), but about 40% of the patients are difficult to maintain the curative effect, showing the rapid reappearance of the receded fear memory. The key to the occurrence and treatment difficulties of PTSD may be the key to the difficulty of occurrence and treatment. Therefore, it is of great significance to study the neurobiological mechanism of the decline and retention of fear. At present, there is not enough study on the process of consolidation and expression relative to the conditioned fear, and many problems remain to be clarified.
Fear receding is that the conditional stimulus (conditioned stimulus, CS) is presented repeatedly without unconditional stimulation (unconditioned stimulus, US, such as the foot shock), and the conditioned fear response gradually diminishes or disappears. It does not erase the previously formed CS-US connection memory, but forms a new memory of CS-no US. A large number of studies have shown that the medial prefrontal cortex (mPFC) in the submarginal zone (infralimbic cortex, IL) participates in the fading process of conditioned fear, which plays an important role in the retention and retention, and is the place for the long-term retention of receding memory. But what is the mechanism of the long-term retention of the memory in the IL region? Which material is the material reference? It is not clear that the retreat process is not yet clear.
On the basis of previous studies, we choose the conditioned fear model and the fear elimination model, the comprehensive behavior, the morphology and the Western-Blot method to explore the role and mechanism of the IL region in the retreat and retention of the rat, in order to find the best "time window" and the key elements of the conditioned fear regression, and to explore the sound conditions in the IL region. The mechanism of sexual fear extinction plays a role in improving the therapeutic effect of PTSD.
Objective: To explore the dynamic changes of NR2B, BDNF and Caspase-3 in the submarginal area of rats after conditioned fear extinction training.
Methods: 75 SD rats were randomly divided into blank control group (group Cont, no treatment), the control group (group N-EXE, non regression training) and 24h subsiding group (EXE group). The conditioned fear model was established by the sound and foot shock paired stimulation. After the fear established 24h, the regression training was carried out, and the stiffness behavior of the rats after the regression training was compared. Changes and changes of NR2B, BDNF and Caspase-3 in IL area.
Result:
1. the changes of the regression and retention of the rats in each group:
After the retreat training, the decline and retention of the rats in the subsiding group increased gradually (fear gradually decreased). After the third and 7 days of the retreat training, the decline of the 24h subsided group was better than that of the control group (P0.01).
2. the changes of NR2B and BDNF immunoreactive substances in IL region of each group.
(1) the content of NR2B and BDNF in the IL area of the control group and the 24h subsided group increased gradually, and increased rapidly on the day 1,3., the highest in the seventh day, but all lower than that in the blank control group (P0.01).
(2) the regression control group and the 24h regression group were compared: there was no significant difference between the two groups in first days; on the 3,7 day, the difference between the two groups was significant, and the 24h regression group was significantly higher than that of the control group (P0.05).
3. the changes of mPFC NR2B and BDNF protein contents in rats of each group:
In the regression control group and the 24h regression group, the IL area NR2B, BDNF protein content increased gradually at the day 1,3,7 day after the regression, and the 1,3 day increased rapidly. The decrease of the control group was basically restored to normal at seventh days. At every time point, the protein content of NR2B and BDNF in the control group was significantly lower than that in the 24h regression group and the blank control group.
4. the changes of Caspase-3 immunoreactive substance in IL region of rats in each group.
Compared with the blank control group, (1) the quality of Caspase-3 immunoreactive substance in the IL region of the control group and the 24h regression group was significantly higher than that in the blank control group. The quality of the caspase-3 immunoreactive substance in the 24h subsiding group was close to the blank control group at the seventh day after the retreat training; (2) the regression control group and 24h subsided with the time of elimination training. The content of Caspase-3 decreased gradually, and the amount of Caspase-3 in the seventh day 24h subsided group basically recovered to normal, and (3) at each time point, the caspase-3 amount in the 24h subsiding group was significantly lower than that of the control group (P0.05).
Conclusion:
The results of 1. behavioral study showed that in the week after the fear elimination training, the regressed control group and the 24h regression group were gradually improved (the conditioned fear improved gradually), and the 24h subsided group was better than the regression control group. It suggested that the retreat training was beneficial to the decline of fear memory. Therefore, the patients of PTSD should be given a timely and active exposure. Treatment;
2. immunohistochemical and Western Blot results showed that the number of immunoreactive cells and protein levels of NR2B and BDNF in IL area were gradually increased with the improvement of regression results, and the highest in 3-7 Tianda after the retreat training. It was suggested that the 3-7 day after the retreat training may be the key time for the fear to decline and the IL region NR2B and BDNF play an important role in the decline and retention of fear. Effect.
After 3. regression training, the content of Caspase-3 in IL area decreased gradually, and the result of 24h subsiding group was better than that of the control group. It was suggested that caspase-3 may be involved in the plasticity of IL region during the memory retention of fear, but the mechanism needs further study.

【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R33

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