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研究生: 范家豪
Fan, Chia-Hao
論文名稱: 評估TrKB之潛力促效劑於阿茲海默氏症細胞與動物模式之神經保護效果
Evaluation of the neuroprotective effects of potential TrkB agonist in Alzheimer’s disease cell and mouse models
指導教授: 謝秀梅
Hsieh, Hsiu-Mei
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: 2019
畢業學年度: 107
語文別: 中文
論文頁數: 85
中文關鍵詞: 阿茲海默氏症類澱粉蛋白Tau蛋白腦源性神經滋養因子TrkB促效劑7,8-二羥基黃酮
英文關鍵詞: Alzheimer's disease, , BDNF, TrkB, Tau, 7,8-DHF
DOI URL: http://doi.org/10.6345/NTNU201901046
論文種類: 學術論文
相關次數: 點閱:41下載:0
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  • 研究背景 1 阿茲海默症(Alzheimer's disease, AD) 1 BDNF/TrkB 訊息傳遞路徑 2 BDNF/TrKB信號傳遞之神經保護功能 3 AD與BDNF/TrkB路徑之關聯 4 BDNF/TrkB通路與AD之假設機制 5 BDNF之類似物 7 研究目的 9 材料與方法 10 實驗動物 10 SH-SY5Y 細胞株之培養 10 細胞存活率分析(MTT assay) 10 Aβ25-35寡聚體製備 11 以小鼠海馬迴初級細胞培養篩選化合物 11 細胞免疫螢光染色分析(Immunocytochemistry, ICC) 12 立體定位手術(stereotaxie apparatus) 13 曠野實驗(open field test) 14 高架十字迷宮(Elevated Plus Maze, EPM) 14 Y字迷宮實驗(Y maze) 15 莫里斯水迷宮實驗(Morris water maze) 15 西方墨點轉漬法(Western blot) 16 免疫組織化學染色(Immunohistochemistry) 18 統計分析(Statistical analysis) 19 研究結果 21 以SH-SY5Y細胞株測試不同濃度化合物之細胞毒性 21 建立小鼠海馬迴之初級細胞培養作為篩藥平台 22 TrkB促效劑之結構類似物可有效緩解Aβ25-35所造成的細胞損傷 22 7,8-DHF、LMDS-1與Aβ25-35對小鼠之運動能力及焦慮行為並無影響 23 LMDS-1可有效的緩解Aβ25-35寡聚體所造成之短期記憶能力損傷 24 7,8-DHF及LMDS-1減緩Aβ25-35寡聚體所導致長期記憶能力損傷 25 LMDS-1可有效減少海馬迴區域的Aβ堆積 26 7,8-DHF與LMDS-1可有效降低小鼠海馬迴DG之Aβ沉積 26 LMDS-1可藉由活化Ras/MAPK路徑來激活CREB轉錄因子,進而上調BDNF之表現量 27 7,8-DHF與LMDS-1可有效緩解因Aβ25-35所導致小鼠海馬迴DG減少pERK之情況 29 7,8-DHF、LMDS-1可有效的緩解因Aβ25-35所導致小鼠海馬迴CA1減少的pCREB的表現量 29 LMDS-1可有效提升小鼠海馬迴BDNF表現量 30 LMDS-1可透過降低GSK-3β之活性,進而減少Tau的磷酸化 30 7,8-DHF、LMDS-1可有效降低小鼠海馬迴DG區域Tau之磷酸化 31 LMDS-1可有效緩解被Aβ25-35所誘導而降低的突觸素synaptophysin表現量 32 LMDS-1、7,8-DHF及Aβ25-35皆未造成明顯的神經發炎 32 討論 34 圖1、BDNF/TrKB之訊息傳遞 38 圖2、實驗時序 39 圖3、使用MTT分析化合物對於SH-SY5Y細胞株之存活率 40 表1、12種化合物對於SH-SY5Y細胞株之IC50 41 圖4、以Aβ25-35處理之初級細胞培養進行篩藥之結果 44 圖5、行為實驗結果 47 圖6、LMDS-1有效降低小鼠海馬迴Aβ蛋白質表現量 48 圖7、7,8-DHF、LMDS-1可有效的降低小鼠齒狀迴之Aβ表現量 50 圖8、LMDS-1藉由活化Ras/MAPK路徑來激活CREB轉錄因子,進而上調BDNF之表現量 55 圖9、7,8-DHF、LMDS-1可有效的緩解因Aβ25-35所導致小鼠齒狀迴減少的pERK的表現量 57 圖10、7,8-DHF、LMDS-1可有效的緩解因Aβ25-35所導致小鼠海馬迴CA1減少的pCREB的表現量 59 圖11、LMDS-1可有效提升小鼠海馬迴BDNF表現量 61 圖12、LMDS-1透過降低GSK-3β之活性,進而減少Tau的磷酸化 64 圖13、7,8-DHF、LMDS-1可有效的降低小鼠齒狀迴的Tau磷酸化 66 圖14、LMDS-1有效緩解Aβ25-35所誘導降低的synaptophysin表現量 68 圖15、LMDS-1、7,8-DHF及Aβ25-35皆未造成明顯的神經發炎 69 圖16、動物行為實驗後之小鼠肝腎及血清檢驗分析報告 72 表2、初級抗體列表 73 表3、次級抗體列表 74 表4、動物行為實驗之結果統整表 75 表5、西方墨點法分析結果之統整表 76 表6、免疫組織化學染色分析結果之統整表 77 參考資料 78

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