本文選題：腦缺血 + 神經(jīng)發(fā)生　； 參考：《南京醫(yī)科大學(xué)》2009年碩士論文

【摘要】：【研究背景和目的】: 成年哺乳動(dòng)物腦內(nèi)存在著神經(jīng)干細(xì)胞,一定條件下可以發(fā)生分裂、增殖,稱為神經(jīng)發(fā)生(neurogenesis)現(xiàn)象。腦損傷、缺血等病理過程能激發(fā)海馬齒狀回區(qū)神經(jīng)干細(xì)胞發(fā)生分裂、增殖。研究表明短暫全腦缺血后,產(chǎn)生的新生神經(jīng)細(xì)胞發(fā)生沿著一定通路遷移,并與周圍的正常細(xì)胞建立突觸聯(lián)系發(fā)揮部分功能,為中樞神經(jīng)損傷后神經(jīng)元的修復(fù)提供了可能。因此,探索腦缺血后調(diào)控神經(jīng)發(fā)生的細(xì)胞內(nèi)信號通路具有十分重要的意義。 Src是一種鈣離子激活的細(xì)胞內(nèi)非受體酪氨酸激酶,在多種組織細(xì)胞的增殖、分化、遷移中發(fā)揮重要作用。本研究探討其在腦缺血再灌注誘導(dǎo)的大鼠海馬齒狀回區(qū)神經(jīng)發(fā)生中的調(diào)節(jié)作用及其具體機(jī)制。 尼莫地平是一種雙氫吡啶類鈣拮抗劑,因其能選擇性擴(kuò)張腦血管而被廣泛應(yīng)用于缺血性腦損傷等疾病的治療,但是否影響神經(jīng)發(fā)生過程,目前報(bào)道較少。為此,本實(shí)驗(yàn)觀察尼莫地平給藥后對腦缺血海馬齒狀回神經(jīng)發(fā)生的影響,并探討其相關(guān)機(jī)制。 【研究方法】: 1.非受體酪氨酸激酶Src在腦缺血再灌注誘導(dǎo)的大鼠海馬齒狀回區(qū)神經(jīng)發(fā)生中的作用機(jī)制研究: 采用四動(dòng)脈阻斷法誘導(dǎo)大鼠全腦缺血,缺血20 min前腦室內(nèi)注射Src的抑制劑SU6656或ERK的抑制劑U0126;免疫組化Brdu標(biāo)記法檢測腦內(nèi)海馬神經(jīng)發(fā)生;Western Blot法檢測海馬組織磷酸化Src(p-Src)、Src蛋白的表達(dá),磷酸化Raf(p-Raf)、Raf蛋白的表達(dá),磷酸化ERK(p-ERK)、ERK蛋白的表達(dá),磷酸化CREB(p-CREB)、CREB蛋白的表達(dá).尼氏體染色檢測海馬神經(jīng)的細(xì)胞損傷。 2.尼莫地平對腦缺血海馬齒狀回神經(jīng)發(fā)生的影響及其相關(guān)機(jī)制的研究: 采用四動(dòng)脈阻斷法誘導(dǎo)大鼠全腦缺血,缺血20min前靜脈注射尼莫地平或腦室內(nèi)注射細(xì)胞外信號調(diào)節(jié)激酶ERK的抑制劑U0126;免疫組化Brdu標(biāo)記法檢測腦內(nèi)海馬神經(jīng)發(fā)生;Western Blot法檢測海馬組織磷酸化ERK(p-ERK)、ERK蛋白的表達(dá)。 【研究結(jié)果】: 1. Src在腦缺血再灌注24小時(shí)后大鼠齒狀回區(qū)持續(xù)激活;預(yù)先給予Src特異性抑制劑SU6656,能有效的減少再灌注7天后齒狀回Brdu陽性細(xì)胞的數(shù)量,抑制再灌注24小時(shí)、72小時(shí)齒狀回區(qū)Raf蛋白在酪氨酸340/341位點(diǎn)的激活以及ERK、CREB蛋白的磷酸化水平;U0126,ERK通路的特異性抑制劑,同樣抑制了再灌注7天后齒狀回新生細(xì)胞的數(shù)量,下調(diào)了ERK、CREB蛋白磷酸化表達(dá),而不影響Src、Raf的表達(dá)。 2.尼莫地平抑制腦缺血再灌注后海馬部位的神經(jīng)發(fā)生的同時(shí)也抑制了腦缺血再灌注后海馬齒狀回p-ERK的表達(dá);海馬部位神經(jīng)發(fā)生在U0126組與U0126+尼莫地平聯(lián)合給藥組差異無統(tǒng)計(jì)學(xué)意義。 【研究結(jié)論】: 1. Src通過調(diào)節(jié)Raf/ERK/CREB信號通路在腦缺血再灌注后海馬區(qū)神經(jīng)發(fā)生中發(fā)揮正向調(diào)節(jié)作用。 2.尼莫地平顯著抑制腦缺血再灌注后海馬齒狀回的神經(jīng)發(fā)生,其機(jī)制與下調(diào)p-ERK表達(dá)密切相關(guān)。
[Abstract]:Background and objective: there are neural stem cells in the brain of adult mammals, which can divide and proliferate under certain conditions. Brain injury, ischemia and other pathological processes can stimulate the division and proliferation of neural stem cells in dentate gyrus. The results show that after transient global cerebral ischemia, the neonate neurons migrate along a certain pathway, and establish synaptic connections with the surrounding normal cells to play a part of the function, which provides the possibility for the repair of neurons after central nervous system injury. Therefore, it is of great significance to explore the intracellular signal pathway regulating neurogenesis after cerebral ischemia. Src is a calcium activated intracellular tyrosine kinase, which proliferates and differentiates in a variety of histocytes. Migration plays an important role. The aim of this study was to investigate the regulatory effect and its mechanism of neurogenesis in dentate gyrus of rats induced by cerebral ischemia and reperfusion. Nimodipine is a dihydropyridine calcium antagonist, which has been widely used in the treatment of ischemic brain injury due to its selective dilatation of cerebral vessels. Therefore, the effect of nimodipine on the development of dentate gyrus after cerebral ischemia was observed, and the related mechanism was discussed. [study method]: 1. The role of non-receptor tyrosine kinase Src in the neurogenesis of dentate gyrus induced by cerebral ischemia-reperfusion in rats: a four-artery occlusion method was used to induce global cerebral ischemia in rats. SRC inhibitor SU6656 or ERK inhibitor U0126was injected into forebrain for 20 min after ischemia, and the expression of phosphorylated Srcnp-Srcnc-Src protein was detected by immunohistochemical Brdu labeling method, and the expression of RAF protein in hippocampal tissue was detected by immunohistochemical Brdu labeling method. The expression of phosphorylated ERKP-ERK protein and phosphorylated CREBB-CREBN-CREB protein. The neuronal damage of hippocampal nerve was detected by Nissl body staining. 2. Effects of nimodipine on dentate gyrus neurogenesis following cerebral ischemia and its related mechanisms: global cerebral ischemia was induced by four-artery occlusion in rats. Nimodipine was injected intravenously before ischemic 20min or U0126, an inhibitor of extracellular signal regulated kinase (ERK), and the expression of phosphorylated ERK- ERK- ERK- ERK protein in hippocampus was detected by immunohistochemical Brdu labeling method. [research results]: 1. SRC was continuously activated in dentate gyrus of rats 24 hours after cerebral ischemia-reperfusion, and the number of Brdu positive cells in dentate gyrus was reduced after 7 days of reperfusion by pretreatment with SRC-specific inhibitor SU6656. Inhibition of activation of Raf protein in dentate gyrus at site 340 / 341 and phosphorylation level of ERKN CREB protein at site 340 / 341 and specific inhibitor of U0126 ERK pathway at 24 h and 72 h after reperfusion also inhibited the number of nascent cells in dentate gyrus 7 days after reperfusion. The phosphorylation of ERKG CREB protein was down-regulated, but the expression of Srctr Raf was not affected. 2. Nimodipine inhibited the neurogenesis of hippocampus after cerebral ischemia and reperfusion, and also inhibited the expression of p-ERK in dentate gyrus after cerebral ischemia and reperfusion. There was no significant difference between U0126 group and U0126 nimodipine group. [conclusion]: 1. Src plays a positive regulatory role in hippocampal neurogenesis after cerebral ischemia-reperfusion by regulating the Rafr / ERK / CREB signaling pathway. 2. Nimodipine significantly inhibited the neurogenesis of dentate gyrus after cerebral ischemia and reperfusion, and its mechanism was closely related to the down-regulation of p-ERK expression.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R363

【共引文獻(xiàn)】

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