本文選題：創(chuàng)傷后應(yīng)激功能障礙 + 認(rèn)知功能障礙��； 參考：《南京大學(xué)》2017年碩士論文

【摘要】：目的術(shù)中知曉是常見的全身麻醉術(shù)中并發(fā)癥,其可導(dǎo)致患者產(chǎn)生嚴(yán)重的神經(jīng)精神疾病,例如創(chuàng)傷后應(yīng)激功能障礙(Post-traumatic stress disorder,PTSD)。PTSD主要表現(xiàn)為對創(chuàng)傷經(jīng)歷的再體驗(yàn)、逃避創(chuàng)傷相關(guān)事件和人、高應(yīng)激三大核心癥狀,使患者承受沉重的神經(jīng)精神負(fù)擔(dān)。目前PTSD的具體病理生理機(jī)制尚不明確,因此有關(guān)于PTSD的研究屬于當(dāng)前研究熱點(diǎn)。神經(jīng)連接蛋白(Neuroligin-1,NLGN-1)主要表達(dá)于興奮性突觸上,胞內(nèi)可通過突觸后密度蛋白-95(Postsynaptic density protein 95,PSD-95)調(diào)控 N-甲基-D-天冬氨酸受體(N-methyl-D-aspartate receptor,NMDAR),羥基-5-甲基-4-異惡唑丙酸受體(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor,AMPAR)的表達(dá),具有調(diào)控興奮性突觸形成及興奮性突觸信號(hào)傳遞的作用,與大腦認(rèn)知功能有密切聯(lián)系。NLGN-1參與自閉癥、阿爾茲海默病(Alzheimer' s disease,AD)等認(rèn)知障礙相關(guān)疾病的病理生理過程。NLGN1也可能與PTSD的情緒、學(xué)習(xí)障礙有關(guān),但具體機(jī)制未明。氯胺酮(Ketamine,KET)是非競爭性的NMDAR拮抗劑,具有抗焦慮、抗抑郁的作用,有望成為新的PTSD治療藥物。本研究中,以NLGN-1為切入點(diǎn),運(yùn)用動(dòng)物行為學(xué)及分子生物學(xué)探討PTSD可能的病理生理機(jī)制及KET對其的作用機(jī)制,為PTSD的防治提供新的思路。方法雄性Sprague-Dawley大鼠(SD大鼠)250-275 g,通過足底電擊建立PTSD動(dòng)物模型。第一批SD大鼠隨機(jī)均分為2組:對照組(Control組)、PTSD組。分別在建模后第7、14天通過條件性恐懼實(shí)驗(yàn)、水迷宮實(shí)驗(yàn)檢測大鼠的恐懼記憶以及學(xué)習(xí)記憶能力;取前額葉皮層、海馬、杏仁核,進(jìn)行免疫印跡(Western blotting,WB)檢測NLGN-1蛋白水平。第二批SD大鼠隨機(jī)均分為4組:對照+生理鹽水組(Control + NS 組)、對照 +KET 組(Control + KET 組)、PTSD+ 生理鹽水組(PTSD+NS組)、PTSO + KET組(PTSO + KET組)。建模30min后腹腔注射KET 2.5 mg/kg,對照組給予同等體積的生理鹽水,連續(xù)腹腔注射14天。于建模后第14天通過條件性恐懼實(shí)驗(yàn)、水迷宮實(shí)驗(yàn)檢測大鼠的恐懼記憶以及學(xué)習(xí)記憶能力;取海馬,進(jìn)行WB檢測NLGN-1、軸突蛋白-1(Neurexin-1,NRXN-1)、PSD-95、NMDAR-2B、AMPA-GluR1 蛋白表達(dá)水平。結(jié)果第一批實(shí)驗(yàn)中,無論是在建模后第7天,還是第14天,在條件性恐懼實(shí)驗(yàn)中,與Control組相比,PTSD組的僵直反應(yīng)時(shí)間占總時(shí)間百分比均顯著增加(P0.01);在水迷宮實(shí)驗(yàn)訓(xùn)練階段,與Control組相比,訓(xùn)練第2、3、4天PTSD組的逃避潛伏期都明顯延長(P0.05);在水迷宮實(shí)驗(yàn)測試階段,與Control組相比,PTSD組的探索目標(biāo)象限時(shí)間均無顯著性差異(P0.05);在WB檢測中,前額葉皮層、海馬、杏仁核的NLGN-1在實(shí)驗(yàn)第7天并無改變(P0.05);在實(shí)驗(yàn)第14天,與Control組相比,PTSD組的前額葉皮層及杏仁核的NLGN-1蛋白表達(dá)水平無顯著差異,但海馬的NLGN-1蛋白水平表達(dá)升高(P0.05)。第二批實(shí)驗(yàn)中,條件性恐懼實(shí)驗(yàn),與PTSD+NS組相比,PTSD+ KET組的僵直反應(yīng)時(shí)間占總時(shí)間百分比顯著減少(P0.01);水迷宮實(shí)驗(yàn)訓(xùn)練階段,與PTSD+NS組相比,訓(xùn)練第2、4、5天PTSD + KET組的逃避潛伏期都明顯縮短(P0.05);水迷宮實(shí)驗(yàn)測試階段,各組的探索目標(biāo)象限時(shí)間并無顯著性差異(P0.05);在WB檢測中,與PTSD + NS組相比,PTSD + KET組海馬的NLGN-1、NRXN-1、PSD-95、NMDAR-2B、AMPA-GluR1蛋白表達(dá)水平都明顯降低(P0.05)。結(jié)論P(yáng)TSD大鼠存在恐懼記憶的增強(qiáng)及海馬相關(guān)的空間學(xué)習(xí)能力的損傷,同時(shí)伴有海馬的NLGN-1表達(dá)升高。給予KET可明顯改善PTSD大鼠的恐懼記憶及海馬相關(guān)的空間學(xué)習(xí)障礙,同時(shí)降低海馬NLGN-1信號(hào)通路分子表達(dá)。本研究結(jié)果表明KET可能是通過降低NRXN-1-NLGN-1-PSD-95通路表達(dá)而抑制海馬過度興奮性發(fā)揮治療作用的。
[Abstract]:Objective intraoperative knowledge is a common complication in general anesthesia, which can lead to severe neuropsychiatric disorders, such as post traumatic stress dysfunction (Post-traumatic stress disorder, PTSD).PTSD, which is mainly manifested in the re experience of traumatic experience, escaping from traumatic events and people, three core symptoms of high stress, and making the patient bear. At present, the specific pathophysiological mechanism of PTSD is not clear, so the research on PTSD is the current research hotspot. Neuroligin-1 (NLGN-1) is mainly expressed in the excitatory synapse, and the intracellular can be regulated by the postsynaptic density protein -95 (Postsynaptic density protein 95, PSD-95) to regulate the N- a. The expression of -D- aspartic acid receptor (N-methyl-D-aspartate receptor, NMDAR) and hydroxyl -5- methyl -4- isooxazole propionic acid receptor (alpha -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor, AMPAR), which regulates excitatory synaptic formation and excitatory synaptic transmission, is closely associated with cognitive function of the brain. Participation in the pathophysiological processes of cognitive impairment related diseases such as autism, Alzheimer's disease, AD, and.NLGN1 may also be related to the mood of PTSD and learning disabilities, but the specific mechanism is not clear. Ketamine (KET) is a non competitive NMDAR antagonist with anti anxiety and antidepressant effects and is expected to become a new PTSD treatment. In this study, in this study, using NLGN-1 as the breakthrough point, the possible pathophysiological mechanism of PTSD and the mechanism of KET on it were explored by using animal behavior and molecular biology to provide new ideas for the prevention and control of PTSD. Methods male Sprague-Dawley rats (SD rats) were 250-275 g, and the animal model of PTSD was established by foot shock. The first batch of SD rats were random. The 2 groups were divided into 2 groups: the control group (group Control) and the PTSD group. The fear memory and learning memory ability of the rats were detected by the conditioned fear experiment on day 7,14 after the modeling. The prefrontal cortex, hippocampus, amygdala, Western blotting, WB were used to detect the level of NLGN-1 protein. The second batch of SD rats were randomly divided into two groups. 4 groups: control + physiological saline group (Control + NS group), control group +KET (Control + KET group), PTSD+ physiological saline group (PTSD+NS group), PTSO + KET group (PTSO + KET group). After modeling 30min, the abdominal injection was 2.5, the control group was given the same volume of saline, continuous intraperitoneal injection for 14 days. After the modeling, the conditioned fear experiment was passed fourteenth days after modeling, The water maze test was used to detect the memory and learning memory ability of rats; take the hippocampus, detect NLGN-1, -1 (Neurexin-1, NRXN-1), PSD-95, NMDAR-2B, AMPA-GluR1 protein expression by WB. Results in the first batch of experiments, seventh days after modeling, or fourteenth days, compared with the Control group, PTSD, PTSD, PTSD, compared with the Control group, PTSD. In the water maze test training stage, the escape latency of group PTSD was significantly longer than that of group Control (P0.05) in the training stage of the water maze test (P0.05), and there was no significant difference in the quadrant time between the PTSD group and the PTSD group in the water maze test stage, compared with the Control group (P0.05). In the WB test, the NLGN-1 of the prefrontal cortex, the hippocampus and the amygdala did not change at seventh days (P0.05). On the fourteenth day of the experiment, there was no significant difference in the level of NLGN-1 protein expression in the prefrontal cortex and amygdala of the PTSD group, but the level of NLGN-1 protein in the hippocampus increased (P0.05). The second batch of experiments showed that the conditioned fear was true. Compared with the PTSD+NS group, the PTSD+ KET group had a significant reduction in the total time percentage of the total time (P0.01), and the escape latency of the PTSD + KET group was significantly shortened (P0.05) in the training phase of the PTSD+NS group compared with the PTSD+NS group, and there was no significant difference in the quadrant time of the exploration targets in the water maze test stage. (P0.05); in the WB test, the expression level of NLGN-1, NRXN-1, PSD-95, NMDAR-2B, AMPA-GluR1 protein in the hippocampus of PTSD + KET group was significantly lower than that of the PTSD + NS group (P0.05). The fear memory of good PTSD rats and the spatial learning disorders related to the hippocampus also reduce the molecular expression of NLGN-1 signaling pathway in the hippocampus. The results of this study suggest that KET may play a therapeutic role in inhibiting hyperexcitability of the hippocampus by reducing the expression of the NRXN-1-NLGN-1-PSD-95 pathway.

【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R749.5
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本文編號(hào)：1876695