Dynamin家族在阿尔茨海默病模型小鼠脑组织中的表达差异分析

发布时间：2019-06-19 07:12

【摘要】：阿尔茨海默病(Alzheimer's disease, AD)是一种以认知功能障碍和记忆减退为主要临床特征的神经退行性疾病。Dynamin家族是一类GTP酶分子,在突触囊泡的循环中起到了关键作用。研究发现该家族参与了Aβ介导的神经毒性,导致了神经功能障碍及AD早期的认知下降。本研究通过检测不同模型小鼠脑组织中dynamin家族分子的表达水平,对dynamin家族参与AD模型鼠认知功能变化的分子机制进行了研究,对dynamin家族作为筛选和评价AD免疫治疗药物效果的分子标记物的可能性进行了初步探讨。首先,在优化脑组织蛋白质提取方法的基础上,本研究检测了不同发育阶段3×Tg AD模型小鼠的脑组织中dynamin家族的表达水平,与对照组相比：在转录水平上,3周龄和3月龄时,dynamin家族分子表达显著下降,最高达83.7%。而6月龄和8月龄时,只有dynamin1的表达下降显著；在蛋白水平上,8月龄的模型小鼠dynamin1和dynamin2的表达分别下降了62%和52.8%,与同龄对照组小鼠相比差异达到显著。结果表明dynamin1的表达水平与AD模型小鼠病程变化呈负相关,提示它可能参与了AD早期的发生和发展。其次,本研究对免疫治疗与否的转基因模型小鼠脑组织中dynamin家族的表达变化进行了分析：被动免疫治疗的3×Tg AD模型小鼠,与同龄未治疗的模型小鼠相比,dynamin1的mRNA和蛋白水平的表达分别上调了59.3%和72%；主动免疫治疗的PDAPP模型小鼠与同龄未免疫的模型小鼠相比,dynamin1的mRNA和蛋白水平分别上调了176.6%和89.8%,且dynamin1蛋白的表达量达到同龄对照组C57小鼠的水平；而未经过免疫治疗的AD模型小鼠,与同龄C57对照组小鼠相比,dynamin1的蛋白表达显著下调。以上结果表明dynamin1的表达上调与以Aβ为靶标的疫苗或者抗体治疗AD的效果呈正相关,有可能治疗组Aβ含量的减少最终恢复了dynamin1的正常表达。综上所述,dynamin1表达的变化与Ap介导的AD病程的早期发展和AD模型小鼠的免疫治疗效果密切相关,dynamin1可以作为AD发病机制重要的候选基因进行更深入的研究,而dynamin1作为分子标记物在药效学上评估AD免疫治疗的效果上具有潜在的应用价值。
[Abstract]:Alzheimer's disease (Alzheimer's disease, AD) is a neurodegenerative disease characterized by cognitive impairment and memory impairment. Dynamic family is a kind of GTP enzyme molecule, which plays a key role in the circulation of synaptic vesicles. It was found that the family was involved in A 尾-mediated neurotoxicity, which led to neurological dysfunction and early cognitive impairment of AD. In this study, the expression level of dynamin family molecules in brain tissue of different model mice was detected, and the molecular mechanism of dynamin family involved in the changes of cognitive function in AD model mice was studied, and the possibility of dynamin family as a molecular marker to screen and evaluate the effect of AD immunotherapy was discussed. Firstly, on the basis of optimizing the method of protein extraction from brain tissue, the expression level of dynamin family in brain tissue of 3 脳 Tg AD model mice at different developmental stages was detected. Compared with the control group, the expression of dynamin family molecules decreased significantly at 3 weeks and 3 months of age, up to 83.7%. At the age of 6 months and 8 months, only the expression of dynamin1 decreased significantly, and the expression of dynamin1 and dynamin2 in the 8-month-old model mice decreased by 62% and 52.8%, respectively, which was significantly different from that of the control group at the same age. The results showed that the expression level of dynamin1 was negatively correlated with the course of disease in AD model mice, suggesting that it may be involved in the early occurrence and development of AD. Secondly, the expression of dynamin family in brain tissue of transgenic mice treated with immunotherapy was analyzed. The expression of mRNA and protein in 3 脳 Tg AD model mice treated with passive immunotherapy was up-regulated by 59.3% and 72%, respectively, compared with that of untreated mice of the same age. Compared with the unimmunized model mice of the same age, the mRNA and protein levels of dynamin1 in PDAPP model mice were up-regulated by 176.6% and 89.8%, respectively, and the expression of dynamin1 protein was up to the level of C57 mice in the same age control group, while the expression of dynamin1 protein in AD model mice without immunotherapy was significantly lower than that in C57 control mice of the same age. These results suggest that the up-regulation of dynamin1 expression is positively correlated with the effect of A 尾 targeted vaccine or antibody in the treatment of AD, and it is possible that the decrease of A 尾 content in the treatment group will eventually restore the normal expression of dynamin1. In conclusion, the change of dynamin1 expression is closely related to the early development of AD mediated by Ap and the immunotherapeutic effect of AD model mice. Dynamin1 can be used as an important candidate gene for the pathogenesis of AD, and dynamin1 as a molecular marker has potential application value in evaluating the effect of AD immunotherapy in pharmacokinetics.
【学位授予单位】：北京交通大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.16

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