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研究生：	王亮博 Wang, Liang-Bo
論文名稱：	探討小分子C-2對Protein-X促進amyloid beta寡聚合及細胞毒性之研究 Investigation of a small molecule, C-2, for its anti- Protein-X inducing amyloid beta oligomerization and cytotoxicity
指導教授：	林榮耀 Lin, Jung-Yaw 賴韻如 Lai, Yun-Ju
學位類別：	碩士 Master
系所名稱：	生命科學系 Department of Life Science
論文出版年：	2020
畢業學年度：	109
語文別：	英文
論文頁數：	39
中文關鍵詞：	阿茲海默氏症、X蛋白、類澱粉蛋白-β肽
英文關鍵詞：	Alzheimer’s disease (AD), Protein-X, β-amyloid (Aβ)
DOI URL：	http://doi.org/10.6345/NTNU202001707
論文種類：	學術論文
相關次數：	點閱：117 下載：0
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阿茲海默氏症（Alzheimer’s disease, AD）已被視為常見的神經退化性疾病，主要病因為海馬迴的神經元受損，同時也引起認知功能障礙的出現。AD特徵包括由細胞內蛋白tau過度磷酸化所造成的神經纖維糾結（neurofibrillary tangles, NFTs）及細胞外類澱粉蛋白-β肽（β-amyloid peptide, Aβ）聚集形成的類澱粉斑塊（plaques）。目前，我們實驗室已發現，與年幼的小鼠相比，年老的AD模式鼠具有較高表現量的X蛋白。為了更進一步了解X蛋白在AD中的作用機制，我們藉由轉殖入X蛋白的SH-SY5Y細胞株作為本實驗使用的模式細胞。在細胞存活率方面以及西方墨點法的實驗中，從寡聚合物Aβ42中處理的細胞組別可以得知，經X蛋白的給予後會產生更多Aβ的聚集，使得細胞存活率降低，而處理小分子藥物的C-2能有效降低X蛋白所給予出的Aβ，進而達到拯救細胞的效果。因此，我們認為C-2可能透過抑制X蛋白而有效治療AD的小分子藥物。

Alzheimer’s disease (AD) has been considered as a common neurodegenerative disorder that causes hippocampal neurons damages and cognitive dysfunctions. AD features include neurofibrillary tangles (NFTs) caused by hyper-phosphorylation of intracellular tau protein, and extracellular β-amyloid (Aβ) plaques by Aβ aggregation. Our laboratory recently found that Protein-X level and the oligomerization of Aβ were concomitantly increased in aged triple transgenic AD mice (3xTg-AD) [PS1M146V, APPK670M/N671L, and tauP301L]. To further understand the mechanism of Protein-X in AD pathway, we established Protein-X stable-transfected SH-SY5Y cell line as the cell model in this experiment. In cell-based system and western blot analysis, Protein-X enhanced the formation of Aβ aggregation and the cytotoxicity is more than that in the present of Protein-X, implying that this cell-based system could serve as a platform for drug screening. Furthermore, we found that the small molecule, C-2, had the ability decrease the Aβ42 cytotoxicity induced by Protein-X. C-2 could be anticipated to develop a promising therapeutic agent for AD.

Introduction  1
Alzheimer’s disease  1
β-amyloid (Aβ) formation  2
The relationship between Aβ and neuronal death  2
AD animal model, triple transgenic mice, (3xTg-AD)  3
Research aims  3


Materials and Methods  4
Materials  4
Cell culture  5
Cell viability assay  5
1 Preparation of Aβ42 for Cell viability assay  6
Western blotting  6
1 Preparation of cell lysates  6
2 Quantification of protein concentration  6
Detemination of Aβ42 oligomerization in the cell-free system by Western blot analysis  7
1 Preparation of the cell-free protein sample  7
2 Aβ42 oligomerization in the cell-free system by Western blot analysis  7
Animals  8
1 Morris water maze test  8
2 Y maze task  8
3 Novel object recognition experiment  9
4 Statistical analysis  9


Results  10
Protein-X enhanced the formation of Aβ42 aggregation with senescence in the brain of 3xTg-AD mice  10
Effects of the small molecule, C-2 on Aβ42 cytotoxicity that enhanced by Protein-X  10
C-2 reduced Aβ42 oligomerization induced by Protein-X in the cell-free system  11
Treatment with the small molecule, C-2, downregulated cell apoptosis markers induced by Aβ42  12
Treatment with the small molecule, C-2, downregulated tau hyper-phosphorylation markers induced by Aβ42  12
C-2 upregulated tau hyper-phosphorylation regulator, GSK-3β  13
C-2 improves the learning and memory deficits of 3xTg-AD mice  13
C-2 treatment ameliorates the behavior on the novel object recognition task in 3xTg-AD mice  14
C-2 treatment significantly reduced memory behaviors in the Y maze task  14


Discussion  15
References  18
Figures  22
                                

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2025/08/05

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