卡路里限制对阿尔茨海默病转基因小鼠神经行为学及脑蛋白组的影响研究

发布时间：2018-04-18 09:19

本文选题：阿尔茨海默病 + 卡路里限制　；参考：《暨南大学》2017年硕士论文

【摘要】：目的:阿尔茨海默病(Alzheimer’s disease,AD)俗称老年痴呆症,是最为常见的一种神经退行性疾病。随着人口老龄化的到来,AD发病率呈急剧上升的趋势。卡路里限制属于饮食限制的一种,但它不会引起营养失调,在AD的众多干预措施中,卡路里限制正越来越显示出其在改善或减轻神经退行性变化方面的强大作用。本文旨在探究卡路里限制对AD转基因小鼠脑蛋白组的影响,进一步为AD的防治提供科学依据。方法:本文选用3月龄的三转基因AD(3xTg-AD)小鼠和野生型(WT)小鼠作为实验小鼠,以40%的卡路里限制(calorie restriction,CR)饮食处理其3个月,同时以随意采食(ad libitum,AL)的3x Tg-AD小鼠和WT小鼠作为对照。首先,通过旷场实验、黑白箱实验和高架十字迷宫实验探究卡路里限制对3xTg-AD小鼠焦虑行为的影响;通过强迫游泳实验和悬尾实验探究卡路里限制对3xTg-AD小鼠抑郁行为的影响;通过水迷宫实验探究卡路里限制对3xTg-AD小鼠记忆行为的影响。随后,利用荧光差异凝胶电泳(2D-DIGE)结合基质辅助激光解析电离飞行时间质谱(MADLD-TOF-MS)技术研究卡路里限制对小鼠海马组织蛋白质组的影响。最后,利用Western blot对差异蛋白进行验证。结果:行为学结果表明,卡路里限制改善了3x Tg-AD小鼠的焦虑、抑郁和空间记忆损伤。蛋白质组学结果表明,在卡路里限制处理的WT小鼠和随意采食的WT小鼠中有11个差异蛋白的表达;在随意采食的3xTg-AD小鼠和随意采食的WT小鼠中有30个差异蛋白的表达;在卡路里限制处理的3x Tg-AD小鼠和随意采食的3x Tg-AD小鼠中有13个差异蛋白的表达。生物信息学分析显示,差异蛋白主要参与代谢、细胞凋亡、细胞间信号传导等进程,且有多个表达异常的蛋白已有研究报道与AD的发生发展密切相关,其中突触相关蛋白synapsin-2(SYN2)和伴侣分子14-3-3 protein epsilon(14-3-3E)经Western blot验证与2D-DIGE结果一致。结论:综上结果表明,卡路里限制改善了3x Tg-AD小鼠的焦虑、抑郁和空间记忆损伤,其中SYN2和14-3-3E等分子的表达改变可能在卡路里限制改善AD转基因小鼠行为学异常中发挥重要作用。
[Abstract]:Objective: Alzheimer's disease (AD), commonly known as Alzheimer's disease, is the most common neurodegenerative disease.With the coming of population aging, the incidence of AD increased sharply.Calorie restriction is one of the dietary restrictions, but it does not cause nutritional disorders. Among the many AD interventions, calorie restriction is increasingly showing its powerful role in improving or mitigating neurodegenerative changes.The purpose of this study was to explore the effect of calorie restriction on brain proteome of AD transgenic mice and to provide scientific basis for AD prevention and treatment.Methods: the 3-month-old and wild-type mice were treated with calorie restriction CR-40% calorie restriction diet for 3 months, and the 3x Tg-AD mice and WT mice fed at random were used as control.Firstly, the effects of calorie restriction on anxiety behavior in 3xTg-AD mice were investigated by open field experiment, black-and-white box experiment and elevated cross maze test, and the effects of calorie restriction on depression behavior in 3xTg-AD mice were investigated by forced swimming test and tail suspension experiment.The effect of calorie restriction on memory behavior of 3xTg-AD mice was investigated by water maze experiment.Then the effects of calorie restriction on proteome in hippocampus of mice were studied by means of fluorescence differential gel electrophoresis (FDGE) 2D-DIGE and MADLD-TOF-MSM (Matrix-assisted laser desorption ionization time of flight mass spectrometry).Finally, the differential protein was verified by Western blot.Results: behavioral results showed that calorie restriction improved anxiety, depression and spatial memory impairment in 3x Tg-AD mice.The proteomics results showed that there were 11 differentially expressed proteins in WT mice treated with calorie restriction and WT mice fed at random, and 30 differentially expressed proteins in 3xTg-AD mice and WT mice fed at will.There were 13 differentially expressed proteins in 3x Tg-AD mice treated with calorie restriction and 3x Tg-AD mice fed at random.Bioinformatics analysis shows that differential proteins are mainly involved in the processes of metabolism, apoptosis and intercellular signal transduction, and many proteins with abnormal expression have been reported to be closely related to the occurrence and development of AD.The synapsin-2 SYN2) and the chaperone 14-3-3 protein epsilon 14-3-3E) were confirmed by Western blot to be consistent with the results of 2D-DIGE.Conclusion: calorie restriction improves anxiety, depression and spatial memory impairment in 3x Tg-AD mice.The changes of expression of SYN2 and 14-3-3E may play an important role in calorie restriction to improve behavioral abnormalities in AD transgenic mice.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16

【参考文献】