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研究生:張訓瑜
研究生(外文):shun-yu Chang
論文名稱:芝麻素衍生物 3-bis (3-methoxybenzyl) butane-1,4-diol 抑制類澱粉蛋白引起的神經傷害
論文名稱(外文):Protective effect of a sesamin derivative, 3-bis (3-methoxybenzyl) butane-1, 4-diol on Aβ-stressed PC12 cells
指導教授:侯建維侯建維引用關係
指導教授(外文):Chien-Wei Hou
學位類別:碩士
校院名稱:元培科技大學
系所名稱:生物技術研究所
學門:醫藥衛生學門
學類:其他醫藥衛生學類
論文種類:學術論文
畢業學年度:102
語文別:中文
論文頁數:101
中文關鍵詞:木脂素芝麻素衍生物類澱粉蛋白阿茲海默症乙醯膽鹼
外文關鍵詞:lignansSesamin derivativesAmyloid betaAlzheimer’s diseaseAcetylcholine
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芝麻素是一種具有抗氧化活性的木脂素 (lignans),已知具有神經保護之功能,不過無法通過血腦障壁。本實驗室與生技中心合作,新合成的芝麻素衍生物 (3-bis (3-methoxybenzyl) butane-1,4-diol, BBD),BBD能通過血腦障壁 (Blood Brain Barrier, BBB)。本實驗研究目的將探討,以PC12神經細胞進行體外實驗,探討類澱粉蛋白(Amyloid beta, Aβ) 刺激神經細胞傷害,同時加入BBD,測試神經細胞傷害程度、發炎物質指數、和神經細胞訊號蛋白質表現情形。實驗結果證實BBD具有神經保護功能。在加入BBD後PC12細胞存活率明顯上升、減少鈣離子釋放、且能抑制細胞傷害率、增加乙醯膽鹼 (Acetylcholine) 釋放、中和自由基的產生與降低脂質過氧化物;西方墨點法 (Western blot) 的實驗結果顯示:BBD能抑制訊號蛋白質p-JNK、p-p38與Bax的表現。與藥物rivastigmine比較,結果顯示BBD與rivastigmine皆能抑制 Aβ逆境傷害,甚至在清除自由基試驗中顯現出BBD更為卓越的效果。以上結果顯示BBD能預防Aβ引神經退化,期望在未來研發阿茲海默症新藥,提供另一類的選擇的潛力。
Sesamin is known for neuroprotection for by its antioxidantive and anti-inflammatory properties, but it’s costly and it lacks blood-brain barrier (BBB) activity. In the present study, we examined the neuroprotective effects of sesamin derivatives, 3-bis (3-methoxybenzyl) butane-1,4-diol (BBD) subjected to amyloid beta (Aβ1-42) in PC12 cells.
  Lipid peroxidation and acetylcholine release from the PC12 cells were tested and reactive oxygen species (ROS), cytokine production, prostaglandin E2 (PGE2) and related signaling pathways from Aβ1-42-induced PC12 cells were examined by enzyme-linked immunosorbent assay (ELISA) or Western blot assay, respectively. BBD showed a protective effect when subjected to Aβ1-42 in PC12 cells. Our results showed that BBD significantly increased cell viability and acetylcholine release, reduced cytotoxicity, malondialdehyde (MDA) and calcium release in PC12 cells.
  BBD significantly reduced Aβ-induced c-Jun N-terminal kinases (JNK), modulated p-p38 activated protein kinase (MAPK) and Bax expression in PC12 cells. The neuroprotective effect of BBD on Aβ1-42 stress model was involved with antioxidantive and anti-inflammatory effects. The result would help the development of new CNS drug for protection of Alzheimer’s disease.
封面.......................................................I
縮寫對照表.................................................II
中文摘要..................................................III
Abstract...................................................V
目錄.....................................................VII
圖目錄....................................................IX
表目錄....................................................XII
第一章 緒論.................................................1
1.1、芝麻...................................................1
1.1.2芝麻素生理活性..........................................3
1.1.2.1、抗氧化作用..........................................4
1.1.2.2、脂質代謝影響........................................5
1.1.2.3、預防高血壓..........................................7
1.1.2.4、維生素E代謝.........................................7
1.1.2.5、其他...............................................8
1.1.3 芝麻衍生物3-BIS (3-METHOXYBENZYL) BUTANE-1,4-DIOL....13
1.2 神經細胞之結構簡敘.......................................17
1.2.1 神經細胞之功能簡敘.....................................19
1.2.2神經元訊息傳遞 (Neurotransmitters).....................20
1.2.3乙醯膽鹼(Acetylcholine)...............................22
1.3退化疾病................................................23
1.3.1神經退化疾病...........................................23
1.3.2阿茲海默症............................................25
1.3.3阿茲海默症症狀.........................................27
1.3.4類澱粉蛋白............................................31
1.5 防禦類澱粉蛋白的逆境反應..................................37
1.5.1 億思能(Exelon).......................................37
1.6 研究動機...............................................39
第二章 材料與方法...........................................41
2.1 藥品.................................................41
2.2 儀器.................................................41
2.3 試劑.................................................41
2.3.2抗體.................................................43
2.4 培養 PC12 細胞........................................44
2.4.2 細胞活化.............................................44
2.4.3 細胞冷凍.............................................44
2.4.4 細胞繼代培養..........................................45
2.4.5 細胞計數.............................................45
2.4.6 樣品處理後之細胞上清液.................................45
2.4.7 芝麻素衍生物 Sesamin derivative-BBD 的製備............46
2.5細胞存活率試驗 (MTT REDUCTION ASSAY FOR CELL VIABILITY)..47
2.6細胞毒性試驗 (LACTATE DEHYDROGENASE RELEASE ASSAY FOR CELLS CYTOTOXICITY)............................................48
2.7鈣離子含量之測定 (CALCIUM CONTENT ASSAY).................49
2.8 TBARS 測試............................................50
2.9 清除活性氧化物能力之測定.................................51
2.10 乙醯膽鹼之測定.........................................52
2.11 前列腺素E2含量之測....................................53
2.12細胞內蛋白質之萃取......................................54
2.13 統計分析.............................................56
2.14 實驗流程.............................................57
第三章 結果...............................................58
3.1 BBD對於 AΒ1-42 的逆境下,對於 PC12細胞毒性實驗............58
3.5 BBD對於 AΒ1-42產生的逆境下對 PC12產生ROS實驗..............62
3.10 BBD對於 AΒ1-42 的逆境下,調控PC12乙醯膽鹼含量實驗.........65
3.11 BBD對於 AΒ1-42 的逆境下,調整PC12前列腺素 E2 含量實驗.....67
3.12 BBD對於 AΒ1-42 的逆境下,調整PC12訊息蛋白的影響...........69
第四章 討論................................................71
第五章 結論................................................75
參考文獻...................................................76

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