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研究生: 吳嘉旋
Wu, Chia-Hsuan
論文名稱: 探討黃芩素衍生物於神經細胞及阿茲海默動物模式小鼠之作用
The effect of Baicalein derivatives on cortical neurons and on an Alzheimer’s disease mouse model
指導教授: 林炎壽
Lin, Yenshou
口試委員: 林炎壽
Lin, Yenshou
梁美智
Liang, Mei-Chih
李冠群
Lee, Guan-Chiun
口試日期: 2024/03/27
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: 2024
畢業學年度: 112
語文別: 中文
論文頁數: 67
中文關鍵詞: 阿茲海默症β類澱粉蛋白DiBAC4(3)Fura-2/AM
英文關鍵詞: Alzheimer's disease, amyloid beta, DiBAC4(3), Fura-2/AM
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202400444
論文種類: 學術論文
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  • 第一章 緒論 1 一、 阿茲海默症之β類澱粉蛋白形成 1 二、 大腦皮質之麩胺酸神經傳遞系統 2 三、 β類澱粉蛋白對麩胺酸傳遞影響 2 四、 阿茲海默症模式小鼠J20 3 五、 臨床治療阿茲海默症藥物 4 六、 中草藥黃芩之功效 5 第二章 研究目的 6 第三章 研究材料與實驗方法 7 一、 藥劑 7 二、 小鼠大腦皮質神經細胞初級培養 8 三、 細胞免疫染色 (Immunocytochemistry, ICC) 8 四、 β類澱粉蛋白寡聚體製備及蛋白質斑點印跡分析 (Dot blot assay) 9 五、 DiBAC4(3)膜電位染劑檢測去極化反應 10 六、 Fura-2/AM (Fura-2-acetoxy-methyl-ester) 鈣離子測量 11 七、 蛋白質萃取、西方墨點法 11 八、 基因型檢測 12 九、 實驗動物與藥物給予 13 十、 莫氏水迷津測試 (Morris water maze) 14 十一、 巴恩斯迷宮 (Barnes maze) 14 十二、 新物體辨識 (Novel Object Recognition) 15 十三、 臉頰採血檢測血清中的GOT、GPT 16 十四、 灌流 (Perfusion) 及肝臟、腎臟組織H&E染色 16 十五、 統計分析 17 第四章 研究結果 18 一、 BE3能抑制細胞因Aβ所誘發的神經細胞不正常去極化 18 二、 BE3能改善NMDA誘導神經細胞不正常去極化 18 三、 BE3無法改善AMPA誘導的神經細胞不正常去極化 19 四、 BE3能抑制細胞內鈣離子因Aβ、NMDA及AMPA所誘發的濃度變化 19 五、 BE3無法直接影響NMDA、AMPA受體所誘導的鈣離子流入 21 六、 BE3可降低由Aβ導致之ERK蛋白磷酸化 22 七、 BE3可降低由NMDA導致之ERK蛋白磷酸化 22 八、 BE3不會影響小鼠體重以及一些基本生理參數 23 九、 給予BE3能改善J20小鼠空間學習記憶以及辨識能力 24 (一)、巴恩斯迷宮 24 (二)、莫氏迷津水迷宮 25 (三)、新物體辨識 26 第五章 討論 27 圖表 31 圖一、BE3不影響膜電位染劑之螢光亮度 32 圖二、BE3抑制因Aβ所誘發的神經不正常去極化 34 圖三、BE3能改善NMDA誘導神經細胞不正常去極化 36 圖四、BE3無法改善AMPA誘導的神經細胞不正常去極化 38 圖五、BE3可以抑制細胞內鈣離子的濃度變化 40 圖六、BE3在第五分鐘不影響離子型麩胺酸接受器NMDA、AMPA receptor 42 圖七、BE3可降低由Aβ導致之ERK蛋白磷酸化 44 圖八、BE3可降低由NMDA導致之ERK蛋白磷酸化 46 圖九、小鼠基因型檢測以及給予BE3後的體重變化 48 圖十、BE3不影響小鼠GOT、GPT以及組織型態 50 圖十一、給予BE3能改善J20小鼠於巴恩斯迷宮之空間學習記憶能力 52 圖十二、給予BE3能改善J20小鼠於莫氏迷津水迷宮之空間學習記憶能力 54 圖十三、給予BE3能改善J20小鼠於新物體辨識之辨識認知能力 56 參考文獻 57 附錄 62 附錄一、神經細胞免疫染色 63 附錄二、β類澱粉蛋白寡聚體製備之形式 65 附錄三、黃芩素 (BE) 及黃芩素衍生物 (BE3) 之結構 67

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