β微管蛋白在创伤性脑损伤大鼠脑组织中的表达规律

发布时间：2019-06-04 12:59

【摘要】：在临床中创伤性脑损伤(traumatic brain injury TBI）是神经外科常见多发疾病，由其导致的病死率、残障率在各类创伤性损伤中比例最高。TBI后患者近、远期易并发癫痫、认知功能障碍等并发症，严重影响患者生存质量及生活质量。由于对TBI后的神经细胞内生化改变缺乏全面认识，因此选择对其有针对性的治疗一直是困扰临床的难点问题。其认识理论、治疗方法、疗效至今仍没有获得突破性进展。大量研究表明微管蛋白（tubulin）家族不但与细胞分裂、增殖、凋亡、肿瘤的发生发展密切相关，还与许多神经系疾病以及学习记忆、认知功能密切相关。β-tubulin广泛表达于多种细胞的胞质中，具有一定的组织和细胞表达特异性，β-tubulin在正常脑组织中表达量高。但国内外尚无TBI后β-tubulin量变引起神经元损伤与认知功能改变之间关系的研究报道。作者拟采用建立大鼠TBI实验模型研究TBI后脑组织中β-tubulin分子的变化规律。目的：探究创伤性脑损伤神经损伤的机理，初步了解细胞骨架蛋白成分之一的β-微管蛋白与创伤性脑损伤之间的关系以及损伤后不同时间的表达量和变化趋势。模拟中度创伤性脑损伤制作TBI大鼠模型，旨在从蛋白表达变化水平角度研究TBI后神经元损伤发生的分子机制，期望通过动物实验提高临床诊治创伤性脑损伤的水平，探索有效脑保护措施的理论依据。方法：选用共70只健康SD大鼠，清洁等级，雄性，体重（220±20g）。动物编号后采用随机法划分为正常对照组（10只）和TBI模型组（60只）；运用头颅瞬间侧向旋转致伤装置制作大鼠TBI模型。按创伤后动物处死时间点划分为6h、12h、24h、72h、7d、14d6个亚组（每组10只）。在各规定时间点处死动物模型，灌注固定脑组织，采集脑组织制作石蜡切片，采用免疫组织化学（immunohistochemistry，IHC）技术检测β-微管蛋白在大鼠海马和皮层组织的分布情况，研究其在脑损伤后脑组织不同部位的表达量及变化规律。将阳性表达部位图片应用图像处理软件转化为灰度值，统计其灰度值，实验数据用均数±标准差（x±SD）表示，应用SPSS17.0统计软件包进行数据分析，采用u检验，设定统计学差异显著水平为P0.05。结果：免疫组化检测结果显示在大鼠海马区、齿状回及大脑皮层下神经细胞中β-tubulin广泛分布在的胞浆、轴突中。对照组的β-tubulin阳性表达量较TBI模型组表达量高，海马区表达量高于皮层区。创伤后不同时间点表达量不同，在TBI后6h模型组β-tubulin表达阳性量少，为最低值（p 0.05），随着创伤时间的推移延长，其表达量逐渐增加。在皮层组织中恢复相对较快，至7d时表达量与正常对照组基本相同。与正常对照组差异比较无统计学意义（p0.05）。但在海马组织区在损伤后14d组β-tubulin表达量与对照组接近，，与正常对照组比较，差异显著性无统计学意义（p0.05）。结论：在创伤性脑损伤模型脑中，大鼠神经元中β-tubulin表达量早期明显下降，随创伤时间推移β-tubulin表达量逐渐升高。创伤对脑组织的不同部位影响不同，不同区域的神经细胞恢复能力不同，皮层区恢复较海马区快，损伤后7dβ-tubulin基本恢复正常水平。海马区至损伤后14d时β-tubulin表达量基本恢复至对照组水平。推论β-tubulin参与神经损伤与修复的过程，有必要继续进一步深入研究其变化的机理、机制，并进行相应的药物干预研究。
[Abstract]:Traumatic brain injury (TBI) is a common multiple of neurosurgery. The mortality and disability rate are the highest among all kinds of traumatic injury. The near-term and long-term complications of patients with TBI, such as epilepsy, cognitive impairment, and the like, seriously affect the quality of life and quality of life of patients. Because of the lack of comprehensive understanding of the biochemical changes in the nerve cells after TBI, the selection of the targeted therapy has been a difficult problem for clinical. The cognitive theory, the treatment method and the curative effect have not yet achieved breakthrough progress. A large number of studies have shown that the tubuin family not only is closely related to cell division, proliferation, apoptosis, and development of the tumor, but also is closely related to many nerve system diseases and learning and memory and cognitive function. Conclusion-tubelin is widely expressed in the cytoplasm of a plurality of cells, and has certain tissue and cell expression specificity, and the expression level of the antigen-tubelin in the normal brain tissue is high. However, there are no studies on the relationship between the changes of neuronal damage and cognitive function after TBI in the home and abroad. The authors proposed to establish a rat TBI experimental model to study the changes of the P-tubuin molecule in the brain tissue after TBI. Objective: To study the mechanism of the nerve injury of traumatic brain injury, and to understand the relationship between the antigen-microtubule protein and the traumatic brain injury of one of the components of the cellular skeleton protein and the expression and change of the time after the injury. Potential: To study the molecular mechanism of the post-traumatic brain injury of TBI from the level of protein expression change, and to explore the theory of effective brain protective measures in order to improve the level of clinical diagnosis and treatment of traumatic brain injury by animal experiments. Methods:70 healthy SD rats, clean grade, male, body weight (220%2) were selected 0 g). The animal number was divided into the normal control group (10 rats) and the TBI model group (60 rats) by using the random method, and the rats were made of TB using the head instant lateral rotation injury device. I model. The time points were divided into 6 h,12 h,24 h,72 h,7 d,14 d6 subgroups according to the post-traumatic animal sacrifice time point (each group 1 (0). The animal model was sacrificed at the specified time points, the brain tissue was fixed by perfusion, the paraffin section was made from the brain tissue, and the score of the antigen-microtubule protein in the hippocampus and the cortex of the rat was detected by using the immunohistochemistry (IHC) technique. The expression of different parts of brain tissue after brain injury and its change after brain injury The image of the positive expression part is transformed into the gray value by the image processing software, the gray value of the image is counted, the average standard deviation (x-SD) of the average number of the experimental data is used for data analysis, the data analysis is carried out by using the SPSS17.0 statistical software package, and the statistical difference is set to P0 by using the u test. .05. Results: The results of immunohistochemistry showed that in the rat hippocampus, dentate gyrus and the neurons in the cerebral cortex were widely distributed in the cells. The positive expression of P-tubuin in the control group was higher than that of the TBI model group, and the expression of the hippocampal region was higher than that of the TBI model group. It was higher than that in cortex area. The expression was different at different time points after trauma. The expression of antigen-tubiulin in the 6-h model group after TBI was small and the lowest value (p.05), and the expression was prolonged with the time of trauma. The amount gradually increased. The recovery was relatively fast in the cortical tissue, and the amount of expression was similar to that of the normal control at 7 d. The group was basically the same. The difference with the normal control group was not statistically significant (p 0.05). However, in the hippocampus, the expression of e-tubuin was close to the control group in 14 days after the injury, and there was no significant difference in the difference between the control group and the control group (p 0.05). Conclusion: In the brain of traumatic brain injury model, the expression of l-tubuin in the rat's neurons was significantly decreased, and with the time of traumas-tubuin. The effect of trauma on different parts of brain tissue was different, and the recovery of nerve cells in different regions was different. The normal level of the expression in the hippocampus from the hippocampus to the 14-day post-injury level It is necessary to continue to study the mechanism and mechanism of the changes in the process of nerve injury and repair.
【学位授予单位】：延安大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R651.15

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