超声微泡联合人参皂苷Rd对阿尔茨海默症转基因小鼠行为学及海马蛋白组学的影响研究

发布时间：2018-04-23 23:11

本文选题：阿尔茨海默症 + 超声联合微泡　；参考：《深圳大学》2017年硕士论文

【摘要】：阿尔茨海默症(Alzheimer’s disease,AD)是常见的老年痴呆症之一,是进行性发展的致死性神经退行性疾病。病人临床表现为认知和记忆功能不断恶化,日常生活能力进行性减退,伴有神经精神症状和行为障碍。利用药物治疗AD需要面临的首要问题是中枢神经系统内存在的血脑屏障阻挡了大部分药物进入大脑。天然药物人参皂苷Rd(ginsenoside Rd,Rd)具有良好的神经细胞保护作用,但对于AD的治疗也面临血脑屏障的问题。目前,低频聚焦超声联合微泡的方法能够短暂、有效、局部地开放血脑屏障,为脑部药物输送和阿尔茨海默症的治疗开辟了一个崭新途径。基于此,本课题提出采用低频聚焦超声联合微泡作为药物输送手段,探索人参皂苷Rd治疗AD转基因动物模型对其行为学及海马蛋白组学的影响。本研究工作采用8月龄的三转基因(PS1M146V/APPSwe/TauP301L,3xTg-AD)小鼠为研究对象,每隔一天对不同分组的AD小鼠进行治疗并持续六周。治疗结束后,从学习记忆能力、焦虑和抑郁行为的改善多个方面对治疗组动物行为学水平进行评估:采用Morris水迷宫和电跳台行为学检测小鼠学习记忆能力;采用旷场实验和高架十字迷宫检测小鼠焦虑程度;采用了悬尾实验检测小鼠抑郁程度。行为学结果表明,与未治疗的3xTg-AD小鼠相比,经超声微泡联合Rd治疗后的3xTg-AD小鼠的学习记忆能力有显著提高,并减轻3xTg-AD小鼠的焦虑,但不能减轻小鼠的抑郁程度;单纯超声微泡与单纯Rd治疗也能一定程度地改善3xTg-AD小鼠的学习记忆能力,但不能减轻3xTg-AD小鼠的焦虑及抑郁程度。在此基础上,采用荧光差异凝胶电泳(two-dimensional fluorescence difference gel electrophoresis,2D-DIGE)联合质谱技术筛选经治疗后的3xTg-AD小鼠海马区差异表达蛋白,与未治疗的3xTg-AD小鼠对比。发现这些差异蛋白中,记忆相关蛋白UCHL1在单纯Rd和单纯FUSMB治疗后的3xTg-AD小鼠海马组织中均表达显著上调。此外,超声微泡联合Rd能改变UCHL1、PGAM1和SYN1蛋白表达,而这三个蛋白可能是参与超声微泡联合Rd改善3xTg-AD小鼠记忆能力的关键分子。本论文的主要创新点主要有:一、对于具有神经元保护功能的天然药物人参皂苷Rd,采用低频超声联合微泡输送其入脑,探索对AD动物的治疗效果,目前尚无报道;二、本课题采用的3xTg-AD小鼠是目前唯一的能同时表现出β-淀粉样蛋白沉积和Tau相关病理特征的最先进的转基因AD动物模型,是本研究的一大特色;三、本课题采用了多方面的行为学水平评估:学习记忆、焦虑和抑郁水平的评估,能够较全面地评估低频超声联合微泡输送人参皂苷Rd入脑这种方法对AD动物的治疗效果。
[Abstract]:Alzheimer's disease (AD) is one of the most common Alzheimer's disease and a fatal neurodegenerative disease. The clinical manifestations of the patients were the deterioration of cognitive and memory function, the progressive decline of daily living ability, and neuropsychiatric symptoms and behavioral disorders. The primary problem with drug therapy for AD is that the blood-brain barrier in the central nervous system prevents most drugs from entering the brain. Ginsenoside Rd(ginsenoside rdd) has a good neuroprotective effect, but the treatment of AD also faces the problem of blood-brain barrier. At present, the combination of low frequency focused ultrasound and microbubbles can open the blood-brain barrier briefly, effectively and locally, which opens a new way for drug delivery in brain and the treatment of Alzheimer's disease. Based on this, this paper proposed the use of low-frequency focused ultrasound combined with microbubbles as a means of drug delivery, to explore the effects of ginsenoside Rd on the behavior and hippocampal proteomics of AD transgenic animal model. In this study, 8-month-old three-transgenic PS1M146V / APPSwe/ TauP301Ln3xTg-ADmice were used to treat AD mice with different groups every other day for six weeks. After the treatment, the behavioral level of the treatment group was evaluated from the aspects of learning and memory ability, anxiety and depression behavior. The learning and memory ability of mice was measured by Morris water maze and electric jump table behavior. Open field test and elevated cross maze were used to test the anxiety degree of mice, and suspending tail test was used to detect the degree of depression in mice. The behavioral results showed that compared with untreated 3xTg-AD mice, the ability of learning and memory of 3xTg-AD mice treated with ultrasound microbubbles combined with Rd was significantly improved, and the anxiety of 3xTg-AD mice was alleviated, but the degree of depression was not alleviated. The treatment of ultrasound microbubbles and Rd could also improve the learning and memory ability of 3xTg-AD mice to some extent, but could not alleviate the anxiety and depression of 3xTg-AD mice. On this basis, two-dimensional fluorescence difference gel electrophoresis2D-DIGE and mass spectrometry were used to screen differentially expressed proteins in hippocampal area of 3xTg-AD mice after treatment, and the results were compared with those of untreated 3xTg-AD mice. It was found that the expression of memory-associated protein (UCHL1) was significantly up-regulated in hippocampus of 3xTg-AD mice treated with Rd and FUSMB alone. In addition, ultrasound microbubbles combined with Rd can change the expression of UCHL1PGAM1 and SYN1 proteins, which may be the key molecules involved in the improvement of memory ability of 3xTg-AD mice by ultrasound microbubbles combined with Rd. The main innovations of this thesis are as follows: first, for the natural drug ginsenoside Rdwhich has neuron protection function, it is transported into the brain by low-frequency ultrasound combined with microbubble to explore the therapeutic effect of AD animal. The 3xTg-AD mice used in this study are the only advanced transgenic AD animal model which can display 尾 -amyloid deposition and Tau related pathological characteristics at present, which is one of the major characteristics of this study. In this study, we used a variety of behavioral assessment: learning and memory, anxiety and depression, which can comprehensively evaluate the therapeutic effect of low-frequency ultrasound combined with microbubble transport of ginsenoside Rd into the brain for AD animals.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16

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