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研究生: 陳品蓉
Chen, Pin-Jung
論文名稱: 探討中草藥純物質 PN 衍生物作為治療阿茲海默氏症藥物之開發與機轉
The development and mechanism of PN derivatives as therapeutic drugs for Alzheimer’s disease
指導教授: 林炎壽
Lin, Yen-Shou
口試委員: 李冠群
Lee, Guan-Chiun
梁美智
Liang, Mei-Chih
林炎壽
Lin, Yen-Shou
口試日期: 2023/06/20
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: 2023
畢業學年度: 111
語文別: 中文
論文頁數: 62
中文關鍵詞: 阿茲海默症β 類澱粉蛋白麩胺酸接受器DiBAC4(3)中草藥
英文關鍵詞: Alzheimer’s disease, amyloid β, glutamate receptor, DiBAC4(3), herbal medicine
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202300960
論文種類: 學術論文
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  • 第一章 緒論 1 一、 阿茲海默症 1 二、 β 類澱粉蛋白形成及神經毒性 1 三、 β 類澱粉蛋白對麩胺酸傳遞 之 影響 2 四、 β 類澱粉蛋白所介導之訊息傳遞路徑 3 五、 阿茲海默症之模式小鼠 J20 4 六、 臨床治療阿茲海默症藥物 4 第二章 研究動機與目的 6 一、 研究動機與目的 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 7 三、 細胞免疫螢光染色法 (IMMUNOCYTOCHEMISTRY, ICC) 8 四、 細胞存活檢測 9 五、 β 類澱粉蛋白寡聚體製備及蛋白質斑點印漬分析 (DOT BLOT ASSAY) 9 六、 DIBAC4(3)膜電位螢光染劑進行藥物篩選 10 七、 西方墨點法 11 八、 基因型檢測 12 九、 實驗動物與藥物給予 12 十、 莫式水迷津測試 (MORRIS WATER MAZE) 13 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 15 十四、 灌流 (PERFUSION)和肝臟、腎臟組織 H&E 染色 16 十五、 統計分析 16 第四章 研究結果 17 一、 純物質 PN1~PN6 對神經細胞之毒性測試 17 二、 PN4 能改善 Aβ 寡聚體所導致的神經細胞不正常去極化 17 三、 PN4 可 透過 AMPAR 及 NMDAR 改善 Aβ 寡聚體所導致的神經細胞不正常去極化 18 四、 PN4 可降低神經細胞經 Aβ 寡聚體所誘導之 ERK 蛋白磷酸化 19 五、 PN4 可降低神經細胞經 NMDA 所誘導之 ERK 蛋白磷酸化 19 六、 給予 PN4 並不會影響小鼠的體重以及其生理參數 20 (一) 基因型檢測 20 (二) 體重測量 20 (三) 發炎指標- GOT、 GPT 21 (四) 肝臟、腎臟組織切片染色 21 七、 給予 PN4 無法改善 J20 小鼠在巴恩斯迷宮之空間學習記憶 21 八、 給予 PN4 無法改善 J20 小鼠在新物體辨識之辨識記憶 22 九、 給予 PN4 可以改善 J20 小鼠在莫氏水迷津之空間學習記憶 22 第五章、討論 24 參考文獻 28 圖表 34 圖一、 純物質 PN1~PN6 對神經細胞之毒性測試 35 圖二、 PN4 不影響 DIBAC4(3)膜電位螢光染劑之螢光亮度 37 圖三、 PN4 能改善 Aβ寡聚體所導致的神經細胞不正常去極化 39 圖四、 PN4 透過 AMPAR 改善 Aβ寡聚體所導致的神經細胞不正常去極化 41 圖五、 PN4 透過 NMDAR 改善 Aβ寡聚體所導致的神經細胞不正常去極化 43 圖六、 PN4 可降低神經細胞經 Aβ寡聚體所誘導之 ERK 蛋白磷酸化 45 圖七、 PN4 可降低神經細胞經 NMDA 所誘導之 ERK 蛋白磷酸化 47 圖八、小鼠基因型檢測以及給予 PN4 後體重的量測 49 圖九、給予 PN4 不影響小鼠的 GOT、 GPT 以及小鼠的組織型態 51 圖十、給予 PN4 無法改善 J20 小鼠在巴恩斯迷宮之空間學習記憶 53 圖十一、給予 PN4 無法改善 J20 小鼠在新物體辨識之辨識記憶 55 圖十二、給予 PN4 可以改善 J20 小鼠在莫氏水迷宮之空間學習記憶 57 附錄 58 小鼠大腦皮質神經元進行初級培養之純度及特性的確認 58 檢測所製備 類澱粉蛋白寡聚體之形式 58

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