阿茲海默症 (Alzheimer's disease; AD) 是其中一個最普遍型態的老化相關神經退化疾病，而且是老年人癡呆的主要因素。在類澱粉蛋白β串聯假說，類澱粉蛋白β的累積對於產生氧化壓力並造成神經毒性是一個很重要的事件。我們的假說是建立在類澱粉蛋白β的聚集或累積會產生氧化壓力造成DNA受損最後產生神經毒性。所以增加DNA修補的效率和DNA的完整性可以拯救類澱粉蛋白β對於神經細胞所造成的神經毒性。為了測試我們的假說，我們使用慢病毒輔導作用 (Lentivirus transduction) 在大鼠神經皮質初代細胞 (Primary cortical neurons) 大量表現類澱粉蛋白β。從西方墨點法和DHE氧化壓力偵測的結果，在前三天類澱粉蛋白β和氧化壓力是最高的，然後持續下降到第七天。DNA受損的標記，Phosphor-histone 2A (γH2AX) 也是呈現相同的結果。昇醣素類似胜肽 (Glucagon-like peptide-1, GLP-1) 是一個生長因子並且有神經保護的功能，給予之後確實降低類澱粉蛋白β和氧化壓力，但γH2AX並沒有被降低。雖然在本實驗中尚未很明顯地改善γH2AX的情形，但GLP-1在於誘發DNA修補的機制仍然是值得探索的領域，因為在阿茲海默症中DNA受損對於神經細胞的傷害是很重要的一環。

Alzheimer's disease (AD) is one of the common form of age-related neurodegenerative diseases and the leading cause of senile dementia. In the beta-amyloid (Aβ) cascade hypothesis, the aggregation of Aβ is a crucial event that produces amounts of oxidative stress contributed to neurotoxicity. In this study, our hypothesis is that Aβ-peptide aggregation enhances oxidative stress which leads to DNA damage and contributes to neurotoxicity. Therefore, increasing DNA repair efficiency and DNA integrity could rescue neuronal cells from Aβ-induced neuronal death. To test our hypothesis, we utilized lentivirus transduction to overexpress Aβ in rat primary cortical neurons. Results of Western blotting and dihydroethidium (DHE) staining have shown that expression of Aβ reached the peak in the first three days as well as the production of reactive oxygen species (ROS), then both Aβ and ROS levels were decreasing in the following day four to day seven. The DNA damage marker, phosphor-histone 2A (γH2AX), demonstrated that neuronal DNA injury was correlated to both levels of Aβ and ROS. Glucagon-like peptide-1 (GLP-1) is a growth factor which has been proved to have neuroprotective properties. After GLP-1 treatment, the production of Aβ and ROS was reduced, but γH2AX was still remaining in 72 hours. GLP-1 has been proved the effects of decreasing Aβ, inflammation and the improvement of recognition, learning and memory in animal model from previous studies. Although we did not see GLP-1 significantly reducing γH2AX, GLP-1 is still a potential drug involving DNA repair. In Alzheimer’s disease, to elevate DNA repair capability is also a important field to investigate in the future.

目錄
論文審定書 i
摘要 ii
Abstract iii
目錄 v
圖次 vi
縮寫 vii
第一章 緒論 1
1. 阿茲海默症 (Alzheimer's disease; AD) 1
2. Tau蛋白的過度磷酸化假說 (The hypothesis of Tau protein hyperphosphorylation) 1
3. 類澱粉蛋白串聯假說 (The amyloid cascade hypothesis) 2
4. 氧化壓力 (Oxidative stress) 4
5. 昇糖素類似胜肽 (Glucagon-like peptide-1, GLP-1) 5
6. 研究動機 6
第二章 材料與方法 7
1. 實驗材料 7
2. 實驗溶液配製 9
3. 實驗儀器 12
4. 實驗方法 12
第三章 結果 16
1. 慢病毒感染初代皮質神經細胞模式之建立 16
2. GLP-1降低類澱粉蛋白β表現 16
3. GLP-1降低類澱粉蛋白β表現徑而下降氧化壓力 17
4. GLP-1緩慢修復已受損DNA 17
第四章 討論 19
參考文獻 23

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