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研究生:李冠青
研究生(外文):Kuan-chung Lee
論文名稱:食品多醣、甲基化衍生物及降解物之抗氧化活性及對腸腺癌細胞之抑制性
論文名稱(外文):Antioxidation and anti-poliferation of food polysaccharides, and polysachharide methylated derivatives and hydrolysates in Caco-2 cells.
指導教授:賴鳳羲
指導教授(外文):Phoency Lai
學位類別:碩士
校院名稱:靜宜大學
系所名稱:食品營養研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2008/01/
畢業學年度:96
語文別:中文
論文頁數:102
中文關鍵詞:腸腺癌細胞食品多醣抗氧化
外文關鍵詞:Cac0-2food polysaccharidesantioxidation
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本研究主要是探討食品多醣膠質、甲基化衍生物及降解物之分子性質與其體外抗氧化活性及抑制腸腺癌細胞之關係,並歸納出具最佳活性之食品多醣。樣品包括: 果膠類、阿拉伯膠、海藻膠質商品與萃取物、玉米穗軸聚木醣、上述多醣降解物與不同分子量劃分物(>10 k及110 k)、石蓮萃取物、以及海藻多醣甲基化衍生物等。抗氧化試驗結果顯示,未降解之食品多醣膠質與植物萃取多醣清除DPPH自由基、ABTS自由基與超氧陰離子的能力皆會隨濃度增加(110 mg/mL)而增加,其中以玉米穗軸聚木醣(Xyl B)、石蓮多醣(GE)與石蓮熱水萃物(GW) 清除ABTS自由基與超氧陰離子之能力最佳(皆可達到100 %),而清除DPPH自由基之能力以石蓮多醣最高,可達到66.9 %。降解後的多醣劃分物中,以聚木醣大分子劃分物(Xyl>10 k) 清除ABTS自由基效果最佳(達100 %),清除超氧陰離子的能力亦達95 %以上。在抑制腸腺癌細胞實驗方面,在1 mg/mL濃度下的數據顯示綠藻石蓴多醣(U4A)具有最高抑制率(達89.7 %),其次為石蓮多醣(抑制率83.9 %)、小麥麩皮聚葡萄醣大分子劃分物(BG>10K) (80.7 %)、-紅藻膠分子量>10k劃分物(ιB>10 k)抑制率較高(74.6 %)、高甲氧基果膠低分子量酵素降解物(HMP110k) (67.6 %)。紅藻多醣衍生物方面,κ-紅藻膠酸降解(κA20)的抑制率(59.1%)較其他紅藻膠衍生物高。綜合以上結果,玉米穗軸聚木醣、小麥麩皮聚葡萄醣、及石蓮多醣具有極佳清除自由基之抗氧化與抑制腸腺癌細胞之效果,石蓴多醣有最好的抑制腸癌細胞之能力,皆值得深分解析其活性機制。其他具有不等程度之生理活性的多醣膠質與萃取物對保健食品之開發也很重要。
This study was to investigate the relationships between the antioxidation and human Caco-2 colon adenocarcinoma cell inhibition of food polysaccharide gums, their methylated derivatives and hydrolysates, in order to conclude the food polysaccharides of best bioactivities. The samples examined included: pectins, Arabic gum, commercial gums and extracts from various algae, corncob xylans, the hydrolysates and large and small molecular fractions of the above samples, Graptopetalum paraguayens extracts, and methylated derivatives of red algal polysaccharides. The results of antioxidation showed that the antioxidant activities such as DPPH-radical, ABTS-radical and superoxide-anions scavenging abilities increased with the concentration (110 mg/mL) for most of original polysaccharides. Among the samples examined, corncob xylans (Xyl B), G. paraguayens polysaccharide (GE) and hot-water extract (GW) showed the greatest ability in scavenging ABTS radicals and superoxide anions (both reach 100%, where GE exhibited the greatest ability in chelating DPPH radicals (66.9%). For the fractions of polysaccharide hydrolysates, the large molecular fractions of xylan hydrolysates (Xyl >10 k) displayed the best effect on removing ABTS radicals (up to 100%) and superoxide anions (>95%). In the case of Caco 2 cell inhibition experiments, the results at 1 mg/mL concentration indicated that Ulva arasakii polysaccharides revealed the greatest inhibition efficiency (89.7%), followed by GE (83.9%), the large molecular fraction of wheat bran glucans (BG>10 k) (80.7%), large molecular fraction of -carrageenan (B>10k) (74.6%), low molecular fraction of high-methoxy pectin hydrolysates by enzyme (HMP1-10k) (67.6%). For the derivatives of red algal polysaccharides, the inhibition effectiveness of -carrageenan acid hydrolysates (A20) (59.1%) was higher than the others. In conclusion, corncob xylans, wheat bran glucans, and GE showed excellent effects in both antioxidation and Caco-2 cell inhibition. Ulva polysaccharide exhibited the best Caco-2 cell inhibition activity. It is worthy to further investigate the bioactivity mechanisms of these samples. The other polysaccharide gums and extracts with various degrees of physiological activities are also important for developing novel healthy foods.
中文摘要……………………………………………………………………………Ι
英文摘要……………………………………………………………………………Π
目錄…………………………………………………………………………………Ⅳ
表目錄………………………………………………………………………………Ⅶ
圖目錄………………………………………………………………………………Ⅷ
名詞縮寫……………………………………………………………………………Ⅸ
壹、前言……………………………………………………………………………1
貳、文獻整理………………………………………………………………………2
ㄧ、食品多醣之定義及特性………………………………………………………2
1.含阿拉伯糖、鼠李糖及半乳糖之多醣
1.1果膠(Pectin)…………………………………………………………………2
1.2阿拉伯膠(Gum arabic)……………………………………………………2
2.水溶性半纖維素
2.1聚木醣(xylan)………………………………………………………………2
2.2聚葡萄糖(β-glucan) ………………………………………………………3
3.酸性海藻多醣
3.1.紅藻膠(carrageenan) ……………………………………………………3
4.綠藻石蓴(Ulva arasakii)多醣……………………………………………3
5.石蓮多醣…………………………………………………………………………3
二、食品多醣之生理活性…………………………………………………………4
1.具生理活性的食品多醣…………………………………………………………4
1.1具抗腫瘤活性之食品多醣的結構、分子特性……………………………5
1.2食品多醣抗腫瘤的可能作用機制……………………………………………6
2.具抗氧化活性的食品多醣………………………………………………………8
三、食品多醣降解作用
1.腸道降解作用機制………………………………………………………………9
2.食品多醣在腸道的功能特性…………………………………………………10
四、抗氧化作用
1.活性氧與自由基………………………………………………………………12
2.抗氧化作用機制………………………………………………………………14
3.抗氧化劑的種類………………………………………………………………17
五、腸腺癌細胞
1. 腸腺癌產生的原因…………………………………………………………22
2. 腸腺癌細胞的特性……………………………………………………………22
3. 細胞活性測定的方法…………………………………………………………23
4. 具有抑制腸腺癌細胞之食品及其機制………………………………………24
參、實驗設計及材料與方法
一、實驗設計………………………………………………………………………25
二、實驗材料
1.材料來源………………………………………………………………………26
2.細胞株…………………………………………………………………………27
3.藥品……………………………………………………………………………27
4.設備……………………………………………………………………………28
三、實驗項目及實驗方法
1. 多醣降解物製備與劃分……………………………………………………29
2. 抗氧化分析
2.1.清除DPPH自由基之效力 …………………………………………………34
2.2螯合亞鐵離子 ………………………………………………………………35
2.3還原力測試 …………………………………………………………………36
2.4清除ABTS自由基離子之效力………………………………………………37
2.5清除超氧陰離子效力 ………………………………………………………39
3.大腸腺癌細胞抑制率分析
3.1.藥品配製 ……………………………………………………………………41
3.2.細胞培養 ……………………………………………………………………42
3.3.細胞存活率分析(MTT assay)……………………………………………44
4.統計分析 ………………………………………………………………………45
肆、結果與討論
ㄧ、食品多醣膠質與植物萃取多醣對腸腺癌細胞的抑制作用
1. 植物萃取多醣對腸腺癌細胞的抑制作用……………………………………46
2. 海藻多醣對腸腺癌細胞的抑制作用…………………………………………49
3. ι-,κ-,λ-紅藻膠與洋菜膠甲基化衍生物對腸腺癌細胞的抑制作用 ……52
4. ι-與κ-紅藻膠(κ car B)及其酸降解物對腸腺癌細胞的抑制作用 ……55
5. 高甲氧基果膠、低甲氧基果膠與阿拉伯膠對腸腺癌細胞的抑制作用……58
二、食品多醣膠質與植物萃取多醣抗氧化活性
1. 清除DPPH自由基能力之影響 ………………………………………………61
2. 清除ABTS自由基的能力之影響……………………………………………65
3. 清除超氧陰離子的能力之影響 ……………………………………………69
4. 還原力之影響 ………………………………………………………………75
5. 螯合亞鐵離子的能力之影響 ………………………………………………76
伍、結論……………………………………………………………………………80
陸、參考文獻………………………………………………………………………82
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