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研究生:黃柏璋
研究生(外文):HUANG,PO-CHANG
論文名稱:杏鮑菇子實體對以Aβ誘導阿茲海默症C57BL/6J小鼠改善記憶學習能力之研究
論文名稱(外文):The study of Pleurotus eryngii fruiting body to ameliorate memory and learning ability in Aβ-induced Alzheimer's disease C57BL/6J mice.
指導教授:梁志弘梁志弘引用關係蔣慎思
指導教授(外文):LIANG,CHIH-HUNGCHIANG,SHEN-SHIH
口試委員:潘子明梁志欽
口試委員(外文):PAN,TZU-MINGLIANG,ZENG-CHIN
口試日期:2016-04-29
學位類別:碩士
校院名稱:東海大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:116
中文關鍵詞:阿茲海默症類澱粉樣胜肽神經纖維糾結杏鮑菇
外文關鍵詞:Alzheimer’s diseaseamyloid β-peptideneurofibrillary tanglesPleurotus eryngii
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阿茲海默症 (Alzheimer's disease) 主要為腦內氧化壓力過高造成類澱粉樣胜肽 (amyloid β-peptide, Aβ) 斑塊堆積及神經纖維糾結 (neurofibrillary tangles, NFTs) 所引發之老年型退化性疾病。杏鮑菇 (Pleurotus eryngii) 子實體含多量之麥角硫因 (ergothioneine)、腺苷 (adenosine) 及多酚 (polyphenol) 等抗氧化活性物質,可減緩因體內氧化壓力所造成之老化現象。故本研究針對杏鮑菇子實體之一般成分及機能性成分進行分析,並探討餵食低、中及高劑量之杏鮑菇子實體粉末對於側腦注射Aβ1-40誘導阿茲海默症之C57BL/6J小鼠學習與認知功能之改善效果。研究結果顯示,一般成分分析之水分、乾物重、粗灰分、粗脂質、粗蛋白及碳水化合物含量,分別為8.34 ± 0.14、91.66 ± 0.14、5.75 ± 0.02、1.55 ± 0.01、16.79 ± 0.11及67.60 ± 0.26 %;機能性成分方面,麥角硫因、腺苷、總酚及多醣體含量,分別為1.36 ± 0.05、3.07 ± 0.08、3.58 ± 0.13及223.89 ± 18.39 mg/g (DB)。動物試驗結果顯示,小鼠餵食杏鮑菇子實體粉末六週後,各組間體重增加量及攝食量無明顯差異。血清生化參數及肝腎組織切片結果亦無異常之情形。在腦組織相對重量結果顯示,控制組、褪黑激素及低、中、高劑量組與Aβ組相比均顯著較高 (p<0.05),顯示餵食杏鮑菇子實體粉末可減緩腦部萎縮之情形。在水迷宮參考記憶試驗中,低、中、高劑量組與Aβ組相比,可明顯減少小鼠於泳池中之逃逸時間 (49、80及85%) 及逃逸距離 (53、68及69%) (p<0.05);在水迷宮空間探索試驗中,於目標象限之徘徊時間、穿梭次數、游泳距離、及游泳速率,各處理組與Aβ組相比,均顯著上升 (p<0.05);T字迷宮試驗結果亦顯示,不同劑量樣品組能顯著提升尋獲食物之正確率 (p<0.05);另發現餵食杏鮑菇子實體粉末可透過減少腦中氧化壓力損傷,顯著降低磷酸化τ-蛋白含量 (phosphorylated τ-protein)、Aβ斑塊沉積量、丙二醛 (malondialdehyde) 及蛋白質羰基氧化物 (protein carbonyl) 含量 (p<0.05),降低神經細胞受損。綜上所述,藉由動物行為模式及腦中阿茲海默相關病徵之氧化物含量分析結果,顯示餵食杏鮑菇子實體粉末可改善小鼠記憶學習及認知功能,可應用於開發具延緩與改善阿茲海默症之相關產品。

Alzheimer’s disease (AD) is major caused by highly oxidative stress, which accelerates the amyloid β-peptide (Aβ) plaques and neurofibrillary tangles (NFTs) accumulated in brain. Pleurotus eryngii (PE) has been known as a rich source of antioxidants including ergothioneine, adenosine and polyphenol, which can reduce the aging phenomenon by oxidative stress in body. The aim of this study was to investigate the proximate compositions and bio-functional compounds in PE and evaluate the cognitive impairment of low (LPE), medium (MPE) and high (HPE) dosages of PE by Aβ induced-Alzheimer’s disease model in C57BL/6J mice. In proximate compositions, the contents of moisture, dry matter , crude ash, crude fat, crude protein and carbohydrate were 8.34 ± 0.14、91.66 ± 0.14、5.75 ± 0.02、1.55 ± 0.01、16.79 ± 0.11 and 67.60 ± 0.26%, respectively. Analysis of bio-functional components, the levels of ergothioneine, adenosine, total phenols and polysaccharide were 1.36 ± 0.05、3.07 ± 0.08、3.58 ± 0.13 and 223.89 ± 18.39 mg/g DB, respectively. The results of Alzheimer’s disease model by 6-week feeding of PE was showed that there is no adverse effects on body weight gain, food efficiency, serum biochemical parameters and histopathological examinations of liver and kidney in all dosages. The relative brain weight of Aβ group was significantly lower than that of other groups (p<0.05). It showed that PE could retard the atrophy of brain. In reference memory task, it showed that LPE、MPE and HPE significantly decreased percentage of escape latency (49-85%) and escape distance (53-69%) compared to that of Aβ group (p<0.05). With regards as the results of probe and T maze tasks indicate that PE administration potently ameliorated the memory deficit in mice. By AD pathology index analysis, PE can significantly decrease the contents of phosphorylated τ-protein, depositions of Aβ plaques, values of malondialdehyde and protein carbonyl in brain. These results suggest that PE can attenuate the memory and learning ability in Aβ-induced Alzheimer's disease mice and have great benefit to development as functional food on prophylaxis of AD.

摘要.......................................................................I
Abstract.................................................................III
表次....................................................................VIII
圖次......................................................................IX
第一章 前言.................................................................1
第二章 文獻回顧.............................................................2
壹、失智症 (dementia)......................................................2
貳、阿茲海默症.............................................................4
一、阿茲海默症之發現及病徵..................................................4
二、阿茲海默症之病理及成因..................................................4
三、阿茲海默症之相關影響因子................................................5
四、阿茲海默症之臨床診斷 ..................................................12
五、阿茲海默症之治療......................................................18
叁、杏鮑菇 (Pleurotus eryngii)............................................22
一、杏鮑菇介紹 ..........................................................22
二、杏鮑菇之營養成分......................................................22
三、杏鮑菇之機能性成分....................................................22
肆、褪黑激素 (melatonin) .................................................32
一、褪黑激素之介紹........................................................32
二、褪黑激素抗氧化機制....................................................34
伍、研究目的與實驗架構.....................................................34
第三章 材料與方法...........................................................37
壹、實驗儀器及藥品.........................................................37
一、實驗儀器.............................................................37
二、藥品試劑.............................................................38
三、酵素免疫分析套組......................................................39
四、實驗材料.............................................................39
貳、實驗方法..............................................................40
一、杏鮑菇子實體一般成分分析...............................................40
二、杏鮑菇子實體機能性成分分析.............................................42
三、體內動物實驗模式......................................................44
四、皮爾森相關係數 (Pearson correlation coefficient)......................56
五、統計分析.............................................................56
第四章 結果與討論...........................................................57
壹、杏鮑菇子實體粉末之一般成分分析...........................................57
貳、杏鮑菇子實體粉末之機能性成分分析.........................................59
一、麥角硫因含量..........................................................59
二、腺苷含量.............................................................59
三、總酚含量.............................................................61
四、多醣體含量...........................................................61
叁、杏鮑菇子實體粉末改善阿茲海默症之研究.....................................62
一、Aβ1-40注射手術及餵食杏鮑菇子實體粉末對C57BL/6J小鼠體重及攝食量之影響......62
二、餵食杏鮑菇子實體粉末對C57BL/6J小鼠血清生化參數之影響.....................62
三、餵食杏鮑菇子實體粉末對C57BL/6J小鼠臟器組織相對重量之影響..................66
四、餵食杏鮑菇子實體粉末對C57BL/6J小鼠腦部組織之影響.........................68
五、餵食杏鮑菇子實體粉末對C57BL/6J小鼠肝腎臟組織之影響.......................68
六、餵食杏鮑菇子實體粉末對C57BL/6J小鼠記憶學習能力之影響.....................73
七、餵食杏鮑菇子實體粉末對C57BL/6J小鼠游泳速率之影響.........................82
八、餵食杏鮑菇子實體粉末對C57BL/6J小鼠於T字迷宮試驗之影響....................82
九、餵食杏鮑菇子實體粉末對C57BL/6J小鼠腦中磷酸化τ-蛋白之影響..................85
十、餵食杏鮑菇子實體粉末對C57BL/6J小鼠腦中丙二醛濃度之影響....................85
十一、餵食杏鮑菇子實體粉末對C57BL/6J小鼠腦中蛋白質羰基氧化物濃度之影響.........88
十二、餵食杏鮑菇子實體粉末對C57BL/6J小鼠腦中Aβ斑塊沉積量之影響................90
十三、餵食杏鮑菇子實體粉末與腦中阿茲海默症病徵及行為能力之相關係數.............93
第五章 結論................................................................96
第六章 參考文獻............................................................99



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