臺灣博碩士論文加值系統

阿茲海默症為最常見的一種神經退化性疾病，過去研究指出血液
和腦部膽固醇代謝異常為阿茲海默症的危險因子之一，主要原因可能是
體內高膽固醇會增加氧化壓力、加速老化及細胞膜損害等造成腦神經細
胞損傷，進而引發神經退化性疾病。在動物和臨床研究上，多酚類物質
已被應用於預防阿茲海默症的發生和改善認知功能障礙，然而有關多酚
類物質對於高膽固醇飲食所造成的神經退化性疾病之動物研究的報告非
常有限。本研究以5%高膽固醇飲食小鼠作為模式，將富含多酚類物質的
紅肉李製成粉末，以2%和5%不同比例添加於飼料中餵養小鼠五個月，
結果顯示5%紅肉李添加可降低由高膽固醇飲食所導致的血清、大腦皮質
和海馬迴膽固醇上升，進而降低BACE 和Cyp46 表現量，使腦中Aβ 和
24-OHC 顯著降低，改善認知功能衰退，進一步對神經退化性疾病達到保
護的作用。

Alzheimer’s disease (AD) is one of the most common forms of
neurodegenerative diseases. It has been shown that the disturbances in brain
and blood cholesterol metabolism are associated with the major pathological
features of AD. High cholesterol contents in the blood and brain could lead to
higher expressions of Aβ protein in the brain and cause damage of brain cells.
Normal aging in animals is accompanied by a decline in cognitive
performance which thought to be due to the long-term effects of oxidative
stress on the neurologic processes. In animal and clinical studies, polyphenols
have been used to prevent the incidence of AD. The purpose of this study was
to observe the effect of the polyphenol-rich oriental plums on the cognitive
functions and on the cerebral neurodegeneration-related protein expressions in
the mice fed 5% cholesterol-enriched diet.
In this study, 5% cholesterol-enriched diet mice were treated with 2% and
5% oriental plum powder for 5 months. The cognitive functions were assessed
using Morris water maze. mRNA expression of Cyp46 was determined by
RT-PCR. Expressions of pathology protein of AD, beta-amyloid (Aβ) and
BACE were assessed by Western blotting. Results showed that consuming the
diet with 5% oriental plum powder significantly decreased plasma and brain
cholesterol, the mRNA expression of Cyp46, the protein of BACE in the brain,
Aβ protein and 24-OHC. In conclusion, consuming 5% oriental plum powder
in diet can improve cognitive function of C57BL/6 mice fed with 5%
cholesterol-enriched diet and may further prevent neurodegenerative diseases.

中文摘要 ....................................................................................................... II
Abstract ......................................................................................................... III
致謝 .............................................................................................................. IV
縮寫表 ........................................................................................................... V
【表目錄】 .................................................................................................. XI
【圖目錄】 ................................................................................................. XII
【附錄】 .................................................................................................... XIII
第一章、緒論 .............................................................................................. 14
第二章、文獻回顧 ...................................................................................... 16
第一節、阿茲海默症 (Alzheimer’s disease, AD) ............................... 16
一、臨床與病理特徵 .................................................................. 16
二、氧化壓力 (oxidative stress) ................................................. 17
三、全球盛行率 .......................................................................... 18
四、台灣盛行率 .......................................................................... 18
第二節、血液膽固醇與神經退化性疾病 ........................................... 19
一、動物模式研究 ...................................................................... 19
二、血液中膽固醇與認知功能障礙 ........................................... 20
三、血液中膽固醇與β 澱粉樣蛋白 (Aβ) 的產生 .................... 20
VII
第三節、腦部膽固醇代謝 .................................................................. 24
一、腦部膽固醇代謝異常 ........................................................... 24
二、腦部膽固醇與神經退化性疾病 ........................................... 25
第四節、高膽固醇飲食與神經退化性疾病的關聯 ........................... 26
第五節、多酚類 (Polyphenols) .......................................................... 27
一、分類 ...................................................................................... 27
二、花青素 (anthocyanins) ......................................................... 28
三、多酚類與神經退化性疾病 ................................................... 28
第六節、紅肉李 (Oriental plum) ........................................................ 30
第七節、實驗假說與目的 .................................................................. 31
第三章、實驗材料與方法 .......................................................................... 32
第一節、實驗動物與實驗設計 ........................................................... 32
第二節、紅肉李粉的製備 .................................................................. 34
一、實驗方法 .............................................................................. 34
第三節、實驗飼料配製 ...................................................................... 35
第四節、紅肉李粉總醣類含量 ........................................................... 36
一、原理 ...................................................................................... 36
二、藥品配置 .............................................................................. 36
三、實驗步驟 .............................................................................. 36
VIII
第五節、 紅肉李粉膳食纖維(Dietary fiber)萃取 ............................. 38
一、實驗步驟： .......................................................................... 38
二、計算公式： .......................................................................... 39
第六節、莫氏水迷宮實驗 (Morris Water Maze)................................ 40
一、實驗流程-空間學習與記憶能力試驗 .................................. 40
二、探索試驗 (probe trial) ......................................................... 41
第七節、樣本採集與前處理 .............................................................. 42
第八節、血液樣品分析方法 .............................................................. 43
一、血漿總膽固醇 (cholesterol) ................................................. 43
二、血清TBARS (thiobarbituric acid reactive substances) ......... 43
第九節、腦部組織檢測項目 .............................................................. 44
一、腦部膽固醇 .......................................................................... 44
二、 大腦皮質與海馬迴Cyp46 mRNA 的表現量 ................... 45
三、大腦皮質與海馬迴24-OHC 的含量 ................................... 46
四、大腦皮質與海馬迴Aβ 和BACE 蛋白質表現量 ................ 48
第十節、統計分析方法 ...................................................................... 52
第四章、結果 .............................................................................................. 53
第一節、紅肉李總醣類及總膳食纖維含量 ....................................... 53
第二節、小鼠體重變化 ...................................................................... 53
IX
第三節、小鼠血膽固醇濃度的變化 ................................................... 53
第四節、小鼠大腦皮質和海馬迴的膽固醇濃度 ............................... 54
第五節、血中膽固醇和腦部膽固醇的相關性 ................................... 54
第六節、血清TBARS ........................................................................ 54
第七節、小鼠認知功能的結果 ........................................................... 55
第八節、大腦皮質和海馬迴Cyp46 的mRNA 表現量 ..................... 55
第九節、大腦皮質和海馬迴24-OHC 的含量 ................................... 56
第十節、大腦皮質和海馬迴中BACE 蛋白的表現量 ....................... 56
第十一節、大腦皮質和海馬迴中Aβ 蛋白的表現量 ......................... 56
第五章、討論 .............................................................................................. 57
第一節、紅肉李總膳食纖維含量 ....................................................... 57
第二節、紅肉李總醣含量和小鼠體重變化 ....................................... 58
第三節、小鼠血清膽固醇 .................................................................. 59
第四節、小鼠腦部膽固醇 .................................................................. 60
第五節、血清膽固醇和腦部膽固醇之相關性 ................................... 62
第六節、高膽固醇飲食和Aβ 的相關性 ............................................ 63
第七節、高膽固醇飲食對認知功能的影響 ....................................... 65
第八節、紅肉李對高膽固醇飲食的影響 ........................................... 67
第九節、紅肉李給予對高膽固醇飲食小鼠認知功能的影響 ............ 68
X
第六章、結論 .............................................................................................. 69
第七章、參考文獻 ...................................................................................... 70

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