胰淀素在正常和痴呆小鼠脑内的分布和表达变化

发布时间：2018-01-20 21:01

本文关键词： 胰淀素老年斑 β-淀粉样蛋白阿尔茨海默病　出处：《重庆医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:系统研究胰淀素在正常和阿尔茨海默病(Alzheimer’s disease,AD)痴呆模型小鼠脑内的分布和表达情况,以及与老年斑的关系,以弄清胰淀素是否参与AD的发病。方法:取5月、9月、12月及以上雄性APP/PS1双转基因小鼠(2×TgAD)和野生型小鼠(WT),每组5只,采用免疫荧光染色结合激光共聚焦技术检测胰淀素在小鼠脑内的分布和表达情况,胰淀素与GFAP、NeuN、老年斑的关系;通过RT-PCR和Western blot(WB)方法检测胰淀素核酸蛋白水平的变化。结果:免疫荧光染色结果显示:胰淀素主要表达于神经元;胰淀素阳性神经元广泛分布于小鼠脑内,但具有区域差异性,强表达于皮质区域,尤其是在颗粒细胞层和锥体细胞层,中等表达于海马各亚区,弱表达于丘脑;在2×TgAD皮质区域胰淀素免疫阳性神经元的数量明显高于WT,而在海马区域则无明显差异;随着年龄的增加,胰淀素阳性神经元的数量明显增多。免疫荧光双标结果显示:随着年龄的增加,胰淀素在2xTgAD小鼠脑内能够聚集形成一些斑块状结构;老年斑的数量逐渐增多;胰淀素和老年斑存在共表达现象。WB显示:胰淀素蛋白在AD小鼠脑内的表达水平高于WT小鼠,且随着年龄的增加,其表达水平逐渐上升。RT-PCR显示:AD小鼠脑内胰淀素mRNA的表达高于WT。结论:胰淀素在不同年龄段正常和AD模型小鼠脑内的分布和表达存在明显差异,这些差异可能与老年斑的形成和AD疾病的发生和发展相关。
[Abstract]:Objective: to study the distribution and expression of amylin in brain of normal and Alzheimer's disease mice. And the relationship with senile plaque to find out whether amylin is involved in AD. Methods: May, September. In December and above, male APP/PS1 double transgenic mice (2 脳 TgAD) and wild type mice (5 mice in each group) were divided into two groups. The distribution and expression of amylin in mouse brain were detected by immunofluorescence staining and laser confocal technique. The level of amylin nucleic acid protein was detected by RT-PCR and Western blotDNA. Results: the results of immunofluorescence staining showed that amylin was mainly expressed in neurons. Amylin positive neurons were widely distributed in the mouse brain, but had regional differences. They were strongly expressed in the cortex, especially in the granular cell layer and pyramidal cell layer, moderately expressed in the hippocampal subareas and weakly expressed in the thalamus. The number of amylin immunoreactive neurons in the cortex of 2 脳 TgAD was significantly higher than that in WT.There was no significant difference in hippocampal area. With the increase of age, the number of amylin positive neurons increased obviously. The results of double immunofluorescence showed that the number of amylin positive neurons increased with age. Amylin could gather in the brain of 2xTgAD mice to form some plaques. The number of senile plaques increased gradually; The co-expression of amylin and senile plaque. WB showed that the expression level of amylin protein in brain of AD mice was higher than that of WT mice, and it increased with age. RT-PCR showed that the expression of amylin mRNA in brain was higher than that of WT.Conclusion:. There were significant differences in the distribution and expression of amylin in the brains of normal and AD mice at different ages. These differences may be related to the formation of senile plaque and the occurrence and development of AD disease.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R749.16

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