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研究生:王彥蘋
研究生(外文):Wang, Yen-Ping
論文名稱:諾麗果汁及乙醇萃取物之抗氧化及抑癌活性
論文名稱(外文):Anti-oxidtive and anti-cancer activities of noni (Morind citrifolia L.)juice and ethnol extracts
指導教授:盧錫祺盧錫祺引用關係
指導教授(外文):Lu, Hsi-Chi
口試委員:蔡正宗江文德蔡政志陳春榮
口試委員(外文):Tsai, Tsun-ChungChiang, Wen-DeeTsai, Cheng-ChihChen, Chun-Jung
口試日期:2012-07-19
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:2012
畢業學年度:100
語文別:中文
論文頁數:130
中文關鍵詞:諾麗抗氧化抑癌
外文關鍵詞:nonianti-oxidtiveanti-cancer
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諾麗(Morinda citrifolia L.)分佈於包括臺灣在內的熱帶環太平洋地域,做為民間保健醫療使用已有兩千年歷史,直至最近才開始系統性科學研究,若干機能性成份分別證實具有不同生理功效,例如諾麗果汁經乙醇處理的多醣體即發現具有抗癌作用。本研究將諾麗果汁進行為期兩個月室溫儲藏,並於每個月取樣後檢測其物化性質(果汁產率、pH值和色澤)、功能性化合物 (總酚類化合物、類黃酮、縮合單寧、東莨萣素及其衍生物和芸香素化合物)和生理活性 (總抗氧化能力和DPPH自由基清除力)。結果顯示,諾麗果汁經儲藏後,部份成分如總酚類化合物、類黃酮類和芸香素化合物之含量與活性即開始損失,部份成分如縮合單寧和東莨萣素含量則會經儲藏而上升。乙醇萃取物以酚硫酸法、還原醣含量與HPLC分析分子量以測定多醣體,發現多醣體的總醣量會因儲藏時間而減少,還原醣含量以儲藏1個月含量較多,而多醣體主產物的分子量在儲放0與1個月時分別為468.61與448.46 KDa,儲存2個月後分析出兩個主產物分別為2739及103.1 KDa。這些隨時間而產生的變化,可能對於抑癌等活性作用產生影響。其次探討諾麗果汁及其乙醇萃取物對人類結腸直腸癌Caco-2細胞的影響。結果顯示,隨著添加濃度的增加,Caco-2細胞存活率均顯著降低,未發酵諾麗果汁於6000 ppm時能使Caco-2細胞存活率低於50%。以流式細胞儀探討細胞凋亡發現諾麗果汁、諾麗果汁乙醇萃取上清液與諾麗果汁乙醇萃取物皆是濃度6000 ppm時,對於Caco-2細胞較能促進早期與晚期凋亡。未來可藉西方轉漬法定量細胞凋亡相關蛋白質表現以探討諾麗抑癌作用之機制。
Noni fruit (Morinda citrifolia L.) which has been used for more than 2000 years in Polynersia as traditional folk medicine is consumed as a food or dietary supplement with purported health benefits. The objective of this study was to investigate the optimal fermentation time, anti-oxidative, and cancer preventive effects of noni juice and ethanol extracts. In order to find the best fermentation time, partial fermented noni juices were prepared from noni fruits stored for three different periods of time: 0, 4, and 8 weeks. The characteristics of noni juice from different fermentation period were assayed by measuring functionalities, including physiological and chemical attributes. The characteristics were analyzed as following (1) changes in the physical properties including total yield, color value, and pH value, (2) changes in the content of functional ingredients including total phenolic compounds, flavonoids, condensed tannins, rutins, scopoletin and its derivatives, (3) changes in the physiological activities including trolox equivalent antioxidant capacity (TEAC) and DPPH free radical scavenging. Experimental results indicated that among three different fermentation periods, noni juice began to reduce some of its functional and physiological attributes such as total phenolic compounds, flavonoids, and rutins as time goes by; while condensed tannins, scopoletin and its derivative were increased over time. To investigate the polysaccharides in noni juice, ethanol extracts of noni juice (noni-ppt) were subjected to phenol–sulphuric acid test, DNS residual sugar assay, and molecular mass determination by HPLC analysis. Experimental results indicated that total sugar of noni-ppt was reduced during fermentation, content of reducing sugar of noni-ppt at the 4th week was higher than other periods, and the molecular mass of two major polysaccharide moieties were 468.61 and 448.46 KDa at 0 and 4 weeks respectively. Addition of noni juice or ethanol extracts to Caco-2 colon cancer cell cultures greatly and dose-dependently decreased cell viability as assayed by MTT. 6000 ppm of unfermented noni juice suppressed more than 50% of Caco-2 cell growth. As showed in flow cytometry, early and late apoptosis were most evident when noni was present at the highest concentration (6000 ppm). Western blot analysis of apoptotsis-related proteins will help to delineate the underlying mechanism.
中文摘要......................................................................1
英文摘要......................................................................2
壹、前言......................................................................3
貳、文獻回顧..................................................................4
2-1 諾麗之簡介................................................................4
2-1-1 名稱.....................................................................4
2-1-2 植物外觀................................................................5
2-1-3產地分佈...................................................................5
2-1-4 品種分類..................................................................9
2-1-5 後熟現象..................................................................9
2-2 諾麗的傳統用途及其加工.......................................................12
2-2-1 傳統用途.................................................................12
2-2-2 諾麗果汁加工.............................................................14
2-3 諾麗的基本成分及其化合物.....................................................19
2-3-1 基本成分性質.............................................................19
2-3-2 酚類化合物...............................................................22
2-3-3 其他重要化合物............................................................28
2-3-4 諾麗的指標性物質..........................................................36
2-4 諾麗果之生理功效............................................................40
2-4-1 抗微生物(antimicrobial) .................................................40
2-4-2 抗氧化(antioxidant activity) ............................................40
2-4-3 抗發炎和止痛活性..........................................................45
2-4-4 抗腫瘤及抗癌(anti-tumor and anti-carcinogenic)...........................45
2-5 果膠多醣之生理特性..........................................................46
2-5-1 .具生理活性的果膠多醣.....................................................46
2-5-2 腸胃道降解作用機制........................................................46
2-5-3 果膠多醣在腸胃道的功能特性.................................................49
2-5-4 諾麗乙醇萃取果膠多醣體.....................................................49
2-6 細胞凋亡...................................................................52
2-6-1 細胞凋亡現象的發現及特徵...................................................52
2-6-2 細胞凋亡相關蛋白質........................................................54
2-6-3 分析凋亡細胞的方法........................................................56
2-6-4 細胞凋亡檢測.............................................................56
2-6-5 細胞凋亡相關蛋白檢測......................................................57
參、材料與方法..................................................................58
3-1 實驗架構...................................................................58
3-2 實驗材料...................................................................60
3-2-1 實驗原料.................................................................60
3-2-2分析試藥與試劑............................................................60
3-2-3 人類細胞株...............................................................61
3-2-4 細胞培養基...............................................................61
3-2-5細胞培養其他藥品...........................................................61
3-2-6 細胞凋亡試劑.............................................................61
3-2-7 蛋白質分析試劑............................................................61
3-2-8 設備儀器.................................................................63
3-3實驗方法....................................................................64
3-3-1發酵諾麗果汁之製備.........................................................64
3-3-2 多醣體製備……………….....................................................64
3-3-3 物理性質.................................................................65
3-3-4 功能性化合物含量測定......................................................65
3-3-5 生理活性分析.............................................................67
3-3-6多醣體分析................................................................69
3-3-7 細胞試驗.................................................................71
3-4 統計分析...................................................................73
肆、結果與討論..................................................................74
Part. 1:諾麗果汁發酵實驗........................................................74
4-1物理性質.....................................................................74
4-2功能性化合物含量..............................................................77
4-3 生理活性...................................................................83
Part. 2:乙醇萃取諾麗果汁沉澱物與抑癌實驗..........................................87
4-4 多醣體分析..................................................................87
4-5 諾麗果汁及乙醇萃取物之抑癌實驗.................................................92
伍、結論.......................................................................107
陸、總結.......................................................................108
柒、參考文獻...................................................................109
捌、附錄.......................................................................121

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