Sombati癫痫细胞模型及红藻氨酸致痫大鼠模型海马中高迁移率族蛋白1表达变化的研究
发布时间：2018-11-02 09:53
【摘要】：第一章Sombati癫痫细胞模型中高迁移率族蛋白1表达变化的研究 【目的】研究Sombati癫痫细胞模型中高迁移率族蛋白1（HMGB1）的变化规律，探讨体外培养的癫痫细胞模型中HMGB1的变化。 【方法】新生SD乳鼠，断头后迅速钝性分离出双侧海马，海马神经元体外培养至第9天，将神经元随机分为模型组（无镁细胞外液处理）和对照组（正常细胞外液处理），用Western-blot技术检测海马神经元在接受相应处理12h、24h、72h后HMGB1蛋白的表达变化情况。 【结果】Western-blot显示，在神经元接受相应处理后12h、24h，模型组细胞所含的HMGB1的量都低于对照组(P0.05);在72h，模型组的表达量明显高于对照组(P=0.006,P0.05)。模型组HMGB1的蛋白表达量于造模后12h（0.2477±0.11810）、24h（0.3331±0.21602）、72h（0.8570±0.25280）呈现逐渐增高趋势。 【结论】Sombati癫痫细胞模型中HMGB1的表达呈时间依赖性，痫性发作12h至72h，其表达量随着癫痫发作时间的延长而增高，HMGB1可能在癫痫的病理生理中起重要作用。 第二章红藻氨酸致痫SD大鼠模型海马区HMGB1表达变化的研究 【目的】探讨红藻氨酸致痫大鼠海马区HMGB1表达的变化情况，以了解癫痫大鼠体内海马区HMGB1的表达变化趋势与体外培养的海马癫痫细胞模型HMGB1的表达变化的异同。 【方法】将40只220 240g的SD雄性大鼠随机分为两组，模型组和对照组，模型组和对照组再各随机分为两组，每组10只，分别为24h组、72h组。模型组以红藻氨酸（KA）经侧脑室注入以诱发大鼠癫痫模型，对照组以等体积的生理盐水代替红藻氨酸。造模后分别于24h和72h处死大鼠，用HE染色法了解红藻氨酸致痫大鼠海马神经元的形态改变，用免疫组织化学技术来检测大鼠海马区HMGB1表达的量和位置改变。 【结果】红藻氨酸致痫大鼠海马区神经元排列稀疏、混乱，胞核浓缩，细胞轮廓模糊，，而对照组未观察到类似的形态改变。造模后24h，模型组中HMGB1的含量(0.1398±0.01801)明显低于对照组(0.2154±0.02873)(p=0.000)，而在72h，模型组HMGB1的含量(0.3652±0.08330)则高于对照组(0.2354±0.00836)(p=0.012)；在这两个时间点观察到的HMGB1蛋白均出现在海马神经元胞质中。 【结论】动物实验的结果类似于体外培养的癫痫细胞模型实验的结果，红藻氨酸致痫大鼠成模后72h，海马区HMGB1出现在海马神经元胞质中且表达增高。
[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 卤0.11810), 24h (0.3331 卤0.21602), 72h (0.8570 卤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 卤0.01801) was significantly lower than that in the control group (0.2154 卤0.02873) at 24 h, while the content of HMGB1 in the model group (0.3652 卤0.08330) was higher than that in the control group (0.2354 卤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.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1
[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 卤0.11810), 24h (0.3331 卤0.21602), 72h (0.8570 卤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 卤0.01801) was significantly lower than that in the control group (0.2154 卤0.02873) at 24 h, while the content of HMGB1 in the model group (0.3652 卤0.08330) was higher than that in the control group (0.2354 卤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.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1
【参考文献】
相关期刊论文 前7条
1 姚咏明,盛志勇;高迁移率族蛋白-1在脓毒症发病中的作用与意义[J];解放军医学杂志;2002年09期
2 张立天,姚咏明,陆家齐,鄢小建,于燕;正丁酸钠对脓毒症大鼠高迁移率族蛋白B1表达的拮抗作用[J];解放军医学杂志;2005年10期
3 马玉东;黎明新;孙宝山;宋岩;;高迁移率蛋白-1水平变化与脓毒症患者死亡率的关系[J];沈阳医学院学报;2010年03期
4 杨智勇,王春友,熊炯p
本文编号：2305691
本文链接：https://www.wllwen.com/yixuelunwen/shenjingyixue/2305691.html
最近更新
教材专著
