【摘要】：第一章Sombati癲癇細(xì)胞模型中高遷移率族蛋白1表達(dá)變化的研究 【目的】研究Sombati癲癇細(xì)胞模型中高遷移率族蛋白1（HMGB1）的變化規(guī)律，探討體外培養(yǎng)的癲癇細(xì)胞模型中HMGB1的變化。 【方法】新生SD乳鼠，斷頭后迅速鈍性分離出雙側(cè)海馬，海馬神經(jīng)元體外培養(yǎng)至第9天，將神經(jīng)元隨機分為模型組（無鎂細(xì)胞外液處理）和對照組（正常細(xì)胞外液處理），用Western-blot技術(shù)檢測海馬神經(jīng)元在接受相應(yīng)處理12h、24h、72h后HMGB1蛋白的表達(dá)變化情況。 【結(jié)果】Western-blot顯示，在神經(jīng)元接受相應(yīng)處理后12h、24h，模型組細(xì)胞所含的HMGB1的量都低于對照組(P0.05);在72h，模型組的表達(dá)量明顯高于對照組(P=0.006,P0.05)。模型組HMGB1的蛋白表達(dá)量于造模后12h（0.2477±0.11810）、24h（0.3331±0.21602）、72h（0.8570±0.25280）呈現(xiàn)逐漸增高趨勢。 【結(jié)論】Sombati癲癇細(xì)胞模型中HMGB1的表達(dá)呈時間依賴性，癇性發(fā)作12h至72h，其表達(dá)量隨著癲癇發(fā)作時間的延長而增高，HMGB1可能在癲癇的病理生理中起重要作用。 第二章紅藻氨酸致癇SD大鼠模型海馬區(qū)HMGB1表達(dá)變化的研究 【目的】探討紅藻氨酸致癇大鼠海馬區(qū)HMGB1表達(dá)的變化情況，以了解癲癇大鼠體內(nèi)海馬區(qū)HMGB1的表達(dá)變化趨勢與體外培養(yǎng)的海馬癲癇細(xì)胞模型HMGB1的表達(dá)變化的異同。 【方法】將40只220 240g的SD雄性大鼠隨機分為兩組，模型組和對照組，模型組和對照組再各隨機分為兩組，每組10只，分別為24h組、72h組。模型組以紅藻氨酸（KA）經(jīng)側(cè)腦室注入以誘發(fā)大鼠癲癇模型，對照組以等體積的生理鹽水代替紅藻氨酸。造模后分別于24h和72h處死大鼠，用HE染色法了解紅藻氨酸致癇大鼠海馬神經(jīng)元的形態(tài)改變，用免疫組織化學(xué)技術(shù)來檢測大鼠海馬區(qū)HMGB1表達(dá)的量和位置改變。 【結(jié)果】紅藻氨酸致癇大鼠海馬區(qū)神經(jīng)元排列稀疏、混亂，胞核濃縮，細(xì)胞輪廓模糊，，而對照組未觀察到類似的形態(tài)改變。造模后24h，模型組中HMGB1的含量(0.1398±0.01801)明顯低于對照組(0.2154±0.02873)(p=0.000)，而在72h，模型組HMGB1的含量(0.3652±0.08330)則高于對照組(0.2354±0.00836)(p=0.012)；在這兩個時間點觀察到的HMGB1蛋白均出現(xiàn)在海馬神經(jīng)元胞質(zhì)中。 【結(jié)論】動物實驗的結(jié)果類似于體外培養(yǎng)的癲癇細(xì)胞模型實驗的結(jié)果，紅藻氨酸致癇大鼠成模后72h，海馬區(qū)HMGB1出現(xiàn)在海馬神經(jīng)元胞質(zhì)中且表達(dá)增高。
[Abstract]:Chapter 1 study on the expression of High Mobility Group protein 1 in Sombati Epilepsy Cell Model [objective] to study the changes of high mobility group protein 1 (HMGB1) in Sombati epileptic cell model. To investigate the changes of HMGB1 in the model of epileptic cells cultured in vitro. [methods] Neonatal SD neonatal rats were divided into model group (free of magnesium extracellular solution) and control group (normal extracellular fluid treatment). The hippocampal neurons were cultured in vitro for 9 days. Western-blot technique was used to detect the changes of HMGB1 protein expression in hippocampal neurons after being treated for 12 h, 24 h and 72 h. [results] Western-blot showed that the amount of HMGB1 in the model group was significantly lower than that in the control group at 12 h and 24 h after the corresponding treatment (P0.05), and at 72 h, the expression of HMGB1 in the model group was significantly higher than that in the control group (P0. 006 P 0.05). In the model group, the protein expression of HMGB1 increased gradually at 12h (0.2477 鹵0.11810), 24h (0.3331 鹵0.21602), 72h (0.8570 鹵0.25280). [conclusion] the expression of HMGB1 in Sombati epileptic cell model is time-dependent. The expression of HMGB1 in epileptic seizures increases with the prolongation of seizure time. HMGB1 may play an important role in the pathophysiology of epilepsy. Chapter 2 study on the changes of HMGB1 expression in hippocampus of epileptic SD rats induced by kainic acid [objective] to investigate the changes of HMGB1 expression in hippocampus of kainic acid-induced epileptic rats. To understand the change trend of HMGB1 expression in hippocampus of epileptic rats and the changes of HMGB1 expression in hippocampal epileptic cell model in vitro. [methods] 40 male SD rats of 220 g were randomly divided into two groups. The model group and the control group were randomly divided into two groups, 10 rats in each group, 24 h group and 72 h group respectively. In the model group, kainic acid (KA) was injected through the lateral ventricle to induce epilepsy in rats, while in the control group, the same volume of normal saline was used instead of kainic acid. The rats were killed at 24 h and 72 h, respectively. The morphological changes of hippocampal neurons in epileptic rats induced by kainic acid were studied by HE staining, and the changes of HMGB1 expression in hippocampus were detected by immunohistochemical technique. [results] the hippocampal neurons of kainic acid induced epileptic rats were sparse, disordered, nucleus concentrated and cell contour blurred, but no similar morphological changes were observed in the control group. The content of HMGB1 in the model group (0.1398 鹵0.01801) was significantly lower than that in the control group (0.2154 鹵0.02873) at 24 h, while the content of HMGB1 in the model group (0.3652 鹵0.08330) was higher than that in the control group (0.2354 鹵0.00836) at 72 h (p0.012). At these two time points, the HMGB1 protein was observed in the cytoplasm of hippocampal neurons. [conclusion] the results of animal experiment are similar to those of the model of epileptic cells cultured in vitro. The expression of HMGB1 in hippocampal neurons appears and increases 72 hours after kainic acid induced epilepsy in rats.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R742.1

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