臺灣博碩士論文加值系統

失智症是一種腦部疾病，其症狀有記憶力退化、反應遲緩、理解及表達能力下降、重複說話等，嚴重者將影響個體的日常運作。記憶力退化主要原因為腦神經細胞持續退化或是死亡，使訊息無法順利的傳遞所導致。成年人的突觸具有可塑性，透過學習及環境的刺激可改變神經突的生長，加強突觸的化學訊息傳遞能力，進而形成記憶並鞏固之。因此，如果能促進腦部神經可塑性，理論上將可強化記憶的形成。許多神經滋養因子，如：腦源性神經滋養因子（Brain-derived neurotrophic factor, BDNF）、膠細胞源性神經滋養因子（glial cell line-derived neurotrophic factor, GDNF）、類胰島素生長因子 （insulin-like growth factors, IGF）等，已經被證實可以促進神經細胞可塑性，進而改善記憶。然而，這類大分子蛋白，往往具有體內結構不穩定、半衰期短、不易通過血腦屏障等問題。由於胜肽藥物具有藥用劑量小、副作用低、易於進入細胞內等優點，因此具有開發優勢。本研究之目的是從我們自行設計的四十條胜肽中，篩選出能影響神經可塑性並促進記憶者，作為藥物開發之候選者。我們以細胞免疫螢光染色法篩選出能夠促進初代海馬迴神經突生長之胜肽，再進一步以莫氏水迷津、被動單向抑制逃避試驗、被動逃避跳台試驗及新穎物體辨識能力測試等記憶相關之動物實驗檢測此胜肽對於記憶障礙大鼠學習與記憶之影響。結果顯示胜肽能促進正常大鼠之空間學習，且能改善經東莨菪鹼或年齡誘導之記憶障礙大鼠的學習與記憶能力，未來將進一步探討其作用機轉。

Dementia is the loss of mental functions such as thinking, memory, and reasoning that is severe enough to interfere with a person's daily functioning. Loss of memory is mainly due to continued degeneration or death of neurons and results in the failure of neural signal transmission. Adult synaptic plasticity such as neurite outgrowth, synaptic transmission, and the formation of memories is strengthened through learning and enriched environmental stimulation. Substances that promote brain neural plasticity, theoretically enhance memory formation as well. Many neurotrophic factors, including brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), insulin-like growth factor (IGF) have been shown to promote neural plasticity as well as memory formation. However, such large proteins usually encountered structural instability, shorter half-life, and are unable to cross the blood brain barrier in vivo. Since peptides play a crucial role in the physiological and biochemical functions, peptide drugs now attracted attention for their potential therapeutic use. Compared with small chemical entity drugs, peptide-based drugs possess certain favorable characteristics, including higher potency, higher selectivity, and better safety. In the present study, forty of peptides were designed and synthesized for neuronal growth assay. Peptides which showed significant effect on neurite outgrowth were selected. The effects of these peptides were further examined by Morris water maze, passive one-way inhibitory avoidance test, passive avoidance step-down test and novel object recognition test for their potential roles in learning and memory. Our results indicate that peptides enhanced learning and memory formation in both naïve, scopolamine-induced and age-derived memory decline amnesia animals. The underlying mechanism of these novel peptides will be further investigated in the future.

中文摘要 I
英文摘要 II
目錄 III
圖目錄 VI
第一章、緒論 1
第一節、神經可塑性 1
（一）神經元生長 1
（二）細胞骨架與可塑性之關係 2
第二節、學習與記憶的形成 3
第三節、失智症 3
第四節、促進記憶之藥物 5
（一）神經滋養因子 5
（二）小分子藥物與可塑性/促進記憶之研究 5
第五節、記憶相關動物實驗 6
第六節、記憶缺失動物模式 7
第七節、研究目的 7
第八節、實驗設計 7
第二章、材料與方法 8
第一節、藥物來源及配製 8
第二節、動物來源及飼養 8
第三節、細胞培養 8
第四節、細胞存活率分析 8
第五節、免疫螢光染色 9
第六節、共軛焦顯微鏡 9
第七節、西方墨點法 9
（一）細胞蛋白質萃取 9
（二）西方墨點法 10
第八節、藥物給予方式 10
（一）正常大鼠 10
（二）藥物誘導記憶障礙模式之大鼠 10
（三）正常老化之大鼠 11
第九節、莫氏水迷津 11
第十節、被動單向抑制逃避學習測驗 11
第十一節、被動逃避跳台測驗 12
第十二節、新穎物體辨識能力測驗 12
第十三節、灌流 12
第十四節、免疫組織染色 13
第十五節、統計分析 13
第三章、結果 14
第一節、胜肽影響神經可塑性 14
（一）胜肽能促進海馬迴神經元生長 14
（二）胜肽增加軸突長度與分枝數 14
第二節、胜肽不會影響神經元存活 15
第三節、胜肽對於細胞骨架蛋白之影響 15
第四節、胜肽能促進正常大鼠的空間學習與記憶表現 16
第五節、胜肽可改善經東莨菪鹼誘導的空間學習和記憶障礙 17
第六節、胜肽可改善經東莨菪鹼誘導的被動單向逃避記憶障礙 17
第七節、胜肽可改善經東莨菪鹼誘導的被動逃避跳台之記憶障礙 18
第八節、胜肽可改善經東莨菪鹼誘導的新穎物體辨識能力 18
第九節、胜肽可改善正常老化之大鼠的空間學習和記憶障礙 19
第十節、胜肽可改善正常老化之大鼠的被動單向逃避記憶障礙 20
第十一節、胜肽可增加大鼠生存壽命 20
第十二節、胜肽能穿透細胞膜作用於細胞質及細胞核 21
第十三節、胜肽能對於大鼠腦部神經元之影響 21
第四章、討論 22
第五章、結論與未來展望 24
第六章、圖 25
附錄 47
參考文獻 48

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