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研究生:黃楓盛
研究生(外文):Feng-sheng Huang
論文名稱:發酵香椿水萃物對老鼠體內抗氧化酵素基因表現之影響
論文名稱(外文):Effects of water extracts from fermented Toona sinensis Roem on the expression of antioxidant enzymes in the Sprague-Dawley Rats
指導教授:鍾雲琴
指導教授(外文):Yun-Chin Chung
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:2008/09/
畢業學年度:96
語文別:中文
論文頁數:68
中文關鍵詞:香椿基因表現動物體內
外文關鍵詞:Toona sinensis Roemgene expression in vivo
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香椿(Toona sinensis Roem)為具有特殊芳香氣味的藥食兩用植物,研究指出香椿在人體生理功能調節上有明顯的效果,如抗腫瘤、抗氧化、降低血糖、提昇人類男性精子活動能力等。許多食品原料在發酵後其抗氧化活性明顯增加 (如天貝、納豆、紅酒等),因此本研究旨於探討經厭氧發酵後香椿水萃物對動物體內抗氧化酵素基因表現之影響。將香椿置於密封罐中經厭氧發酵14天後,以熱水萃取30分鐘並冷凍乾燥製成發酵香椿水萃物(WFTS)。以Sprague-Dawely(SD)大鼠為動物實驗模式評估WFTS對體內抗酵素基因表現之影響。實驗分為控制組以及餵食WFTS 高劑量組(0.5 g/Kg BW)及低劑量組(1.0 g/Kg BW),經連續管灌28天,以RT-PCR(Reverse Transcription Polymerase Chain Reaction)評估肝臟中抗氧化酵素基因(catalase、GPx、SOD) 之mRNA表現量,蛋白質表現量則以西方墨點法(Western blot) 檢測。結果顯示,餵食WTFS 1.0g/Kg BW 後SD大鼠肝臟中catalase、GPx、SOD等抗氧化酵素基因之mRNA表現量會明顯增加,但只有catalase的蛋白質表現量達顯著提升(p<0.05),而SOD蛋白量並未受到WFTS的影響,因此推估WFTS可藉提升catalase、GPx、SOD等抗氧化酵素基因的轉錄作用, SOD的轉譯作用受到了干擾,以至於SOD蛋白表現量無增加。
Toona sinensis Roem is one kind of plant with special fragrant, and its leaves are commonly used for food as well as medication. Researches demonstrated that Toona sinensis possesses many biological functions such as anticarcinogen, increasing antioxidant activity, lowering blood sugar, and raising human sperm activity. Fermentation processing enhances antioxidant activities for food materials, for examples of tempeh, natto, and red wine. The objective of this study was to evaluate the effect of the Water extract from fermented toona sinensis(WFTS) on the gene expression levels in Sprague-Dawely (SD) rats. Toona sinensis Roem leaves were canned and fermented for 14 days, and then the water extract from the fermented leaves was obtained by boiling the leaves in water for 30 min, and then the supernatant was freeze-dried. SD rats were allocated with body weight-basis randomization to three groups, each consisting of 8 males, and orally gavaged with 0, 0.5 and 1.0 g/kg body weight of WFTS for 28 days, respectively. The gene expression of antioxidant enzymes (catalase, GPx, SOD)were determented by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and western blotting. The results of this study showed that oral administration of WFTS at a dosage of 1.0 g/Kg BW could increase catalase, GPx and SOD mRNA expression in liver of SD rats. In protein expression, only the catalase was increased significantly (p&lt;0.05) while in SOD, no significant change was noted by WFTS treatments. The present resulst suggested that WFTS may boost transcription for catalase, GPx and SOD; however, SOD translation was inhibited in someway.
圖目錄 IV
表目錄 VI
中文摘要.................................................................................................VII
Abstract VIII
前言 1
第一章、文獻回顧 2
一、香椿 2
(一) 簡介 2
(二) 香椿成分分析 5
(三) 香椿之生理活性 7
二、自由基與抗氧化 8
(一) 自由基的種類 9
(二) 抗氧化物質 12
三、發酵產品之抗氧化活性 17
四、發酵香椿之相關研究 18
第二章、材料與方法 20
一、實驗流程 20
二、實驗材料 21
(一)、動物實驗方面 21
(二)、化學藥品 21
(三)、使用儀器 24
三、實驗方法 25
(一)、樣品來源及配製 25
(二)、動物實驗設計 25
(三)、Total RNA抽取 27
(四)、反轉錄聚合酶鏈鎖反應(Reverse Transcription Polymerase Chain Reaction;RT-PCR)。 27
(五)、臟器處裡 31
(六)、SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) 32
(七)、西方點墨法(Western blotting) 33
第三章、結果與討論 37
一、發酵香椿水萃物對SD大鼠肝臟抗氧化酵素相關基因之mRNA表現之影響。 37
(一)引子設計 37
(二)、抗氧化酵素相關基因之半定量聚合酶鏈鎖反應條件(semi-quantitative PCR)設定 37
(三)、抗氧化酵素之mRNA表現量 49
二、發酵香椿水萃物對SD大鼠肝臟抗氧化酵素基因之蛋白質表現量 54
(一) catalase之蛋白質表現量 54
(二) Cu/Zn-SOD之蛋白質表現量 54
(三) Mn-SOD之蛋白質表現量 54
三、討論 58
第四章、結論 61
參考文獻 63
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