臺灣博碩士論文加值系統

蕃茄是廣泛存在於市面上的一種橘色至紅色的天然蔬果，近年來其主要深紅色成分蕃茄紅素(lycopene)在許多癌症的預防飲食保健上已被廣泛的研究及探討，但對於其扮演預防癌症角色的機制尚未明瞭，因此本研究主要以蕃茄及其製品與蕃茄紅素為研究之主要對象，先從化學檢定探討其抗自由基主作用方向切入，再以生物檢定方式─亦即選取Salmonella Typhimurium TA102[ Histidine mutation : hisG428 (pAQ1 )] 採用Ames test方式，來探討蕃茄及主成分色素其扮演預防癌症的機制。
引用日本學者Okubo所提出的原理公式︰P = K[X]．[Y]．[Z]，使用化學發光儀測量抗氧化物質Y化學發光(Chemiluminescence；CL )的強度P來判斷該抗氧化物質之清除自由基的能力。X為活性氧化物種(reactive oxygen species, ROS )，本研究使用H2O2為活性物種來源，Y為抗氧化劑即為蕃茄試品，Z為自由基接受體( free radical receptor )
，本實驗使用acetaldehyde( MeCHO )。K為常數。化學發光P值( CL count )愈高就代表抗氧化的程度愈高。
本研究探討單獨使用β-胡蘿蔔素、維生素C、蕃茄、蕃茄製品、蕃茄紅素、沒食子酸( gallic acid ) 對抗氧化作用之效果。結果顯示︰在化學檢定下，蕃茄、蕃茄製品及番茄紅素在無自由基接受體的存在下，發光強，抗氧化效果顯著；而β-胡蘿蔔素、維生素C及沒食子酸在無自由基接受體的存在下，發光微弱，抗氧化效果不明顯。生物檢定顯示，清除過氧化氫自由基能力以蕃茄紅素及蕃茄製品的效果最強，其抑制百分比分別為lycopene 97%，β-carotene 48% ，ascorbic acid 24%；濃度劑量反應曲線上亦顯示隨著蕃茄紅素的濃度增加產生回覆突變的菌落數目愈少，求得IC50為0.102 mM。本研究結果綜合之：蕃茄及其製品都具有顯著的抗氧化暨抗變異效果。

Tomatoes are natural food. many searches are interested in the subject of anticancer by healthy diets. Although scientists have focused on the effect of lycopene which is the main dark-red pigment in tomatoes in recent years, the procession how lycopene works is still unknown. Consequently, our study aims to tomatoes, tomato products and lycopene. We have used the chemiluminescence method to observe the ability of decreasing the free radical damage. Afterward we have applied of Ames test by using Salmonella Typhimurium TA102 tester strain〔Histidine mutation : hisG428(pAQ1)〕in order to know the roles of tomatoes, tomato products and lycopene in preventing cancer.
According to Okubo’s formula P = K 〔X〕〔Y〕〔Z〕, we adjudge the free radical scavenging capacity of an antioxidant by P which is measured by a chemiluminescence reader. X is reactive oxygen species(ROS). Hydrogen peroxide (H2O2) is used as the reactive oxygen species in our study. Tomatoes, tomato products and lycopene were treated as the antioxidant Y. Z is a free radical receptor. We choose acetaldehyde to be Z. K is a constant. P signifies the chemilumunescence count ( CL ) and P value shows the degree of antioxidant.
We have compared the antioxidant activities of β-carotene, Vitamin C, tomatoes, tomato products, lycopene and gallic acid. As the result in chemical testing, tomatoes, tomato products and lycopene show high CL without the free radical receptor. They have higher abilities of antioxidation than β-carotene, Vitamin C and gallic acid while the free radical receptor did not exist. In vitro, our result indicates that the free radical scavenging capacity of lycopene and tomato products were better than other samples. Suppression ratio of mutagenecity(%) of lycopene was calculated to be 97, β-carotene was 48, and ascorbic acid was 24, respectively. The revertant colonies decreased as the concentration of lycopene was increasing has shown on the dose-response curve. The IC50 for lycopene was estimated at 0.102mM. As described above, our experimental results showed that tomatoes and tomato products express some useful effects in antimutagenicity and antioxidant.

【目錄】
第一章 前言︰研究目的...............................1
第二章 文獻探討︰研究背景
第一節 自由基的定義...........................3
第二節 自由基在生物體內的作用.................4
第三節 自由基的來源...........................6
第四節 過多自由基造成的傷害...................7
第五節 抗氧化與自由基.........................8
第六節 安姆氏沙門氏菌( Ames test )試驗.......10
第七節 鼠傷寒桿菌變異株(Salmonella typhimurium)
TA102 的選用..........................13
第三章材料與方法
第一節 材料與試劑............................15
第二節 實驗原理與方法........................17
第四章結果
第一節 研究「自由基、抗氧化劑、自由基接受體」
在化學發光儀中之分佈情行..............28
第二節 各種不同濃度gallic acid抗氧化情形.....29
第三節 比較市售蕃茄品種及市售蕃茄加工製品的
抗氧化能力 ........................ 30
第四節 蕃茄紅素與維生素C抗氧化的效果 .......32
第五節 抗突變性分析 ........................33
第五章 討論 ...................................35
第六章 結論與建議 .............................. 38
第七章 參考文獻 .................................40
圖表
圖1 ……………………………………………………………… 46
圖 2 ……………………………………………………………… 47
圖 3 ……………………………………………………………… 48
圖 4 ……………………………………………………………… 49
圖 5 ……………………………………………………………… 50
圖 6 ……………………………………………………………… 51
表1 ……………………………………………………………… 52
圖7 ……………………………………………………………… 52
圖8 ……………………………………………………………… 53
圖9 ……………………………………………………………… 54
圖10 …………………………………………………………… 55
圖11 …………………………………………………………… 56
圖12 …………………………………………………………… 57
圖13 …………………………………………………………… 58
表2 ………………………………………………………………59
表3 ………………………………………………………………60
表4 ………………………………………………………………61
圖14 …………………………………………………………… 61
表5 ………………………………………………………………62
圖15 …………………………………………………………… 62
圖16 …………………………………………………………… 63
圖17 …………………………………………………………… 64
圖18 …………………………………………………………… 65
表 6 ……………………………………………………………66
附錄
附錄一 自由基的種類及其代謝物………………………………………74
附錄二 自由基對生物體分子的傷害和可能引起的病變………………75
附錄三 ROS與RNS細胞內生性來源路徑……………………………… 76
附錄四 自由基的來源和導致的結果………………………………… 77
附錄五 plasmid pAQ1的結構………………………………………‥78
附錄六 α-胡蘿蔔素、β-胡蘿蔔素、蕃茄紅素之化學的結構式 79
附錄七 四等分畫線平板…………………………………………‥80
附錄八 安姆氏試驗( Ames test )之標記試驗( marker test ) … 81
附錄九 標記試驗( Marker Test )紀錄表………………………… 82
附錄十 以Mitomycin C誘導TA102基因性突變…………………… 83
附錄十一 蕃茄及其製品中類胡蘿蔔素之含量…………………… 84
附錄十二 自由基對細胞膜的傷害………………………………… 85
附錄十三 抗氧化劑對致癌物形成的調控點…………………………86

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