發(fā)布時間：2018-04-22 15:31

  本文選題：大腦皮質(zhì)發(fā)育障礙 + mTOR信號通路　； 參考：《西南醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究揭示哺乳動物雷帕霉素靶蛋白(mTOR)的特異性抑制劑雷帕酶素(Rap)是否可以阻斷X射線宮內(nèi)照射孕鼠而引起的大腦皮質(zhì)發(fā)育障礙(DCDs)的形成發(fā)展,為靶向mTOR信號通路治療DCDs引發(fā)的癲癇及認(rèn)知損害提供一定的實驗基礎(chǔ),為更好的治療難治性癲癇(IE)提供實驗依據(jù)。方法:(1).將12只健康SD孕鼠隨機(jī)分為4組,即分別為正常組、模型組、模型組+溶劑組(溶劑組)、雷帕霉素組,每組3只,于大鼠懷孕16天時分別給予溶劑組腹腔注射二甲基亞砜溶劑(DMOS)及雷帕霉素組腹腔注射雷帕霉素藥物處理,于懷孕17天對除正常組以外的三組予以X射線(195c GY)宮內(nèi)照射。取各實驗組宮內(nèi)胚胎期大鼠為實驗對象。(2).各實驗組分別取懷孕18天、20天、22天母鼠1只解剖取得宮內(nèi)胚胎期大鼠頭部,將腦組織浸泡于4%多聚甲醛中固定3天,孕20天、孕22天大鼠行頭顱顱骨剝離,進(jìn)行腦組織取材,每個試驗點取6只胚胎期大鼠,行石蠟包埋,石蠟切片,腦切片厚5μm,每50μm取一片(即每間隔9片取1片),分別取3只胚胎期大鼠腦組織行常規(guī)蘇木精-伊紅染色(HE染色)、硫堇尼氏染色(Nissl染色),觀察大腦皮質(zhì)和海馬結(jié)構(gòu)的病理特點。(3).對HE染色和Nissl染色切片額葉皮層、海馬皮層、海馬區(qū)病理圖片進(jìn)行采集,對各實驗組病理圖大腦皮層的運(yùn)動區(qū)、感覺區(qū)進(jìn)行皮層厚度測量比較。結(jié)果:(1).HE染色:模型組與正常組比較,可見皮層變薄、胼胝體部分缺失、變薄、皮層結(jié)構(gòu)紊亂,分界不清,海馬結(jié)構(gòu)異常,異常結(jié)節(jié),海馬連續(xù)性中斷等病理改變。溶劑組與正常組比較,可見與模型組類似的異常病理結(jié)構(gòu)改變,雷帕霉素組與正常組比較,可見部分組織胼胝體變薄,皮層異常病變,病理改變不及模型組和溶劑組明顯。(2).Nissl染色:模型組與正常組比較,可見皮質(zhì)變薄、胼胝體部分缺失、皮層結(jié)構(gòu)異常,海馬結(jié)構(gòu)異常等病理改變。溶劑組可見具有與模型組類似的病理結(jié)構(gòu)表現(xiàn)。雷帕霉素組與正常組比較,可有部分組織出現(xiàn)病理改變,但病理改變嚴(yán)重程度不及模型組和溶劑組。(3).大腦皮層厚度分析:E18,各實驗組間皮層厚度比較無明顯差異(P0.05)。E20,E22,額葉運(yùn)動皮層區(qū),前肢區(qū)感覺皮層區(qū),海馬-前肢區(qū)運(yùn)動皮層區(qū),海馬-前肢區(qū)感覺皮層區(qū)各部位模型組與正常組比較,皮層厚度變薄(P0.05)。溶劑組與模型組比較,皮層厚度未見明顯差異(P0.05)。雷帕霉素組與正常組皮層厚度比較分析皮層無明顯差異(P0.05)。雷帕霉素組與模型組比較,模型組皮層厚度變薄(P0.05)。雷帕霉素組與溶劑組比較,溶劑組皮層厚度變薄(P0.05)。結(jié)論:(1).X射線宮內(nèi)照射E17孕鼠可導(dǎo)致胚胎鼠出現(xiàn)異常神經(jīng)細(xì)胞群,形成了皮質(zhì)發(fā)育障礙;(2).雷帕霉素可能干預(yù)了X射線宮內(nèi)照射E17胚胎鼠大腦皮質(zhì)發(fā)育障礙的形成;(3).DCDs大鼠大腦皮層厚度的檢測揭示了其腦體積減少的可能是皮層厚度的減少。
[Abstract]:Aim: to investigate whether rapamycin Rapin, a specific inhibitor of rapamycin target protein in mammals, can block the formation and development of cerebral cortex dysplasia induced by intrauterine irradiation in pregnant mice. It provides experimental basis for the treatment of epilepsy and cognitive impairment caused by DCDs by targeting mTOR signaling pathway and provides experimental basis for better treatment of intractable epilepsy. The method is 1: 1. Twelve healthy SD pregnant rats were randomly divided into 4 groups: normal group, model group, solvent group (solvent group, rapamycin group, 3 rats in each group). The rats were given intraperitoneal injection of dimethyl sulfoxide (DMOS) and rapamycin intraperitoneally at the 16th day of pregnancy, and the three groups except normal group were given intrauterine X-ray irradiation at 17 days after pregnancy. The intrauterine embryonic rats in each experimental group were selected as experimental objects. In each experimental group, one female rat was dissected from 18 to 20 days of gestation to obtain the head of the rat in fetal phase, and the brain tissue was immersed in 4% polyformaldehyde for 3 days, then 20 days of gestation and 22 days of gestation were performed, and the brain tissue was taken from the rat. Six embryonic rats were taken from each experimental site and embedded in paraffin wax sections. The thickness of brain slice was 5 渭 m, one slice was taken every 50 渭 m, and three embryonic rat brain tissues were stained with hematoxylin and eosin staining and Nissl staining respectively. The pathological characteristics of cerebral cortex and hippocampal structure were observed. Pathological pictures of frontal cortex, hippocampal cortex and hippocampal area were collected by HE staining and Nissl staining, and the thickness of motor and sensory cortex of cerebral cortex were measured and compared in each experimental group. Results compared with the normal group, the pathological changes such as thinning of cortex, loss of corpus callosum, thinning of corpus callosum, disorder of cortical structure, unclear boundaries, abnormal hippocampal structure, abnormal nodules and continuity of hippocampus were observed in the model group. Compared with the normal group, the solvent group showed similar abnormal pathological changes as the model group, and the rapamycin group showed that the corpus callosum thinned and the abnormal cortical lesion occurred in some tissues compared with the normal group. The pathological changes were not as obvious as those in the model group and solvent group. Compared with the normal group, the cortical thinning, the loss of corpus callosum, the abnormal cortical structure and the abnormal hippocampal structure were observed in the model group. The solvent group showed similar pathological features as the model group. Compared with the normal group, some tissues in rapamycin group showed pathological changes, but the pathological changes were not as serious as those in the model group and solvent group. There was no significant difference in cortical thickness among the experimental groups (P0.05, E20E22, frontal motor cortex, forelimb sensory cortex, hippocampal / forelimb motor cortex, hippocampal / forelimb motor cortex). Compared with the normal group, the cortex thickness of the model group was thinner than that of the normal group. There was no significant difference in cortical thickness between solvent group and model group (P 0.05). Comparison of cortical thickness between rapamycin group and normal group there was no significant difference in cortex (P 0.05). Compared with the model group, the cortex thickness of the model group was thinner than that of the rapamycin group. Compared with solvent group, the cortex thickness of rapamycin group was thinner than that of solvent group (P 0.05). Conclusion the abnormal nerve cell group of the embryonic mice could be induced by intrauterine irradiation of 1: 1 X ray in the pregnant mice of E17, resulting in cortical dysplasia. Rapamycin may have interfered with the formation of cerebral cortical dysplasia in E17 embryonic mice after intrauterine X-ray irradiation. The measurement of cerebral cortex thickness in rats with DCDs revealed that the decrease of cerebral volume may be the decrease of cortical thickness.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R742.1

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