大鼠机械性脑损伤动态变化的蛋白质组学研究

发布时间：2018-11-06 20:25

【摘要】： 目的:建立体外培养神经细胞和大鼠创伤性脑损伤动物模型,研究损伤后培养神经细胞形态学改变和大鼠海马蛋白质组表达情况,寻找差异性生物标志蛋白。方法:选新生24小时内SD大鼠,取海马组织,培养10-14天后,随机分为对照组及损伤后4h、8h、12h、24h和48h组,复制Scott's神经细胞液压冲击损伤模型,进行神经细胞形态学观察。选成年雄性SD大鼠随机分为手术对照组及损伤后4h、8h、12h、24h和48h组,复制Marmarou's落体打击大鼠脑损伤模型,采用双向凝胶电泳技术分析大鼠闭合性脑损伤后不同时间海马中蛋白质组表达的改变。结果:1体外培养神经细胞液压冲击损伤装置稳定性评价:通过记录小室内和有机玻璃管内的压力大小来检测液压冲击损伤压力的大小,结果表明,小室内和有机管内的压力存在一线性关系(Y=4.4763X+0.2986,Y代表有机玻璃管内压力,X代表小室内压力),造成细胞损伤力的大小以小室内的压力为准。有机玻璃管和小室内压力随钟摆打击角度的增加而增大。轻(0.1MPa)、中(0.2MPa)和重度(0.3MPa)液压冲击损伤所需钟摆下落角度分别为13.5°、21.5°和27°。2液压冲击损伤神经细胞形态学改变:液压冲击后,神经细胞甲苯胺蓝染色,正常神经细胞胞核呈蓝色,胞浆染成淡蓝色,损伤组培养神经细胞主要表现为部分神经细胞胞体浓缩,胞核固缩,染色较深,呈深蓝色,多为坏死型损伤细胞,
[Abstract]:Aim: to establish an animal model of cultured nerve cells and traumatic brain injury (TBI) in vitro, to study the morphological changes of cultured neurons and the proteome expression in hippocampus of rats after injury, and to search for different biomarkers. Methods: SD rats within 24 hours were selected and cultured for 10-14 days. The rats were randomly divided into two groups: the control group and the control group. The Scott's nerve cells were induced by hydraulic shock for 24 hours and 48 hours after the injury. The morphology of nerve cells was observed. Adult male SD rats were randomly divided into two groups: the control group and the control group. The rats were divided into two groups at 4 h, 8 h, 12 h and 48 h after injury. The model of brain injury was induced by Marmarou's falling body. Two-dimensional gel electrophoresis was used to analyze the changes of proteome expression in hippocampus of rats after closed brain injury at different time points. Results: 1 Stability evaluation of the hydraulic shock injury device for cultured nerve cells in vitro: the hydraulic impact injury pressure was measured by recording the pressure in the chamber and in the plexiglass tube. There was a linear relationship between the pressure in the chamber and the pressure in the organic tube (YY 4.4763X 0.2986 Y for the pressure in the plexiglass tube and X for the pressure in the chamber), and the damage force of the cells was determined by the pressure of the chamber. The pressure of plexiglass tube and chamber increases with the increase of pendulum angle. The falling angles of pendulum for 0.1MPa, 0.2MPa and 0.3MPa were 13.5 掳, 21.5 掳and 27 掳respectively. The nerve cells were stained with toluidine blue, the nuclei of normal neurons were blue and the cytoplasm was light blue. In the injured group, the nerve cells in the injured group were mainly concentrated in the cell bodies, pyknosis of the nuclei, deep staining and dark blue. Most of them are necrotic injury cells.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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