資料載入中.....
|
請使用永久網址來引用或連結此文件:
http://libir.tmu.edu.tw/handle/987654321/36418
|
| 題名: | 台灣蔬果及藥材之自由基清除與乳癌生長之活性探討 |
| 作者: | 張莘儀 |
| 貢獻者: | 生藥學研究所 |
| 日期: | 2010 |
| 上傳時間: | 2010-10-20 13:06:47 (UTC+8) |
| 摘要: | 台灣處於亞熱帶，四季物種豐富且多樣，尤其水果和天然植物更是台灣不可或缺的資產，已有許多文獻指出，天然植物當中普遍含有大量多酚類化合物， 其中多酚類化合物更被研究發現具有抗老化、抗氧化、預防心血管疾病與清除自由基功能。自由基為帶有不成對電子的原子、分子或離子，生物體常見的自由基為氧自由基 ( Oxygen free radicals )，種類包含氫氧自由基 ( Hydrogen radical ) 等，易導致致癌基因 ( Oncogene ) 活化或抑癌基因 ( Tumor suppressor gene ) 失去功能，進而引發癌症產生。癌症即惡性腫瘤，由於近年來罹患人數與死亡人數持續高攀，已成為世界衛生組織與醫療研究機構關注的焦點，而乳癌目前是台灣女性發生率最高的癌症，其死亡率高居第四位，且發生率與死亡率仍逐年上升，與歐美國家相比，更有年輕化的現象。
本論文收集台灣本土蔬菜9種、水果2種、藥材7種、香料與其他植物3種，總計21種天然植物，並分自由基清除能力與乳癌細胞之生長活性能力測試兩部分加以探討研究：
< 第一部份 >： 21種蔬果及藥材自由基清除之能力
多酚含量測定結果包含：總多酚含量（Gallic acid equivalent，GAE）( μg Gallic acid / mg）以：咖啡 ( GAE = 160.75 ± 4.36 )、百里香 ( GAE = 194.47 ± 4.99 )、紫蘇 ( GAE = 192.41 ± 4.88 )、蔓澤蘭 ( GAE = 227.20 ± 9.33 )、桂花( GAE = 254.74 ± 5.87 )、龍眼花( GAE = 718.78 ± 7.37 ) 和YC-27 ( GAE = 631.28 ± 8.12 ) 之多酚含量較高。總類黃酮含量（Rutin equivalent，RE）( μg Rutin / mg）則以：咖啡 ( RE = 12.75 ± 0.23 )、百里香 ( RE = 24.12 ± 0.98 )、紫蘇 ( RE = 22.98 ± 1.20 )、蔓澤蘭 ( RE = 30.72 ± 1.32 )、龍眼花 ( RE = 17.47 ± 0.52 ) 和YC-27 ( RE = 45.29 ± 2.23 ) 之總類黃酮含量較高。總黃酮醇含量 （ Rutin equivalent，RE ） ( μg Rutin / mg ） 則以：咖啡 ( RE = 30.63 ± 0.57 )、百里香 ( RE = 29.64 ± 0.83 )、山藥 ( RE = 21.99 ± 0.87 )、紫蘇 ( RE = 31.36 ± 1.49 )、蔓澤蘭 ( RE = 26.03 ± 1.19 )、龍眼花 ( RE = 29.40 ± 0.68 ) 和YC-27 ( RE = 86.08 ± 3.52 ) 之總黃酮醇含量較高。
經過一系列抗氧化測試後，發現其中良好抗氧化之萃取物包括：咖啡、百里香、地瓜、原生種山藥、洛神花、紫蘇、蔓澤蘭、桂花、龍眼花和YC-27有顯著清除DPPH自由基能力，咖啡、百里香、紫蘇、蔓澤蘭、桂花和龍眼花有顯著FRAP還原能力，虎斑瓠皮和火龍果皮有顯著螯合亞鐵離子能力，黑胡椒有顯著抑制超氧陰離子能力，苦瓜、洋蔥、百里香、龍眼花和YC-27顯著清除超氧陰離子能力，咖啡、百里香、地瓜、原生種山藥、山藥、洛神花、紫蘇、蔓澤蘭、桂花、龍眼花和YC-27具有顯著抑制肝粒線體脂質過氧化之能力。為了進一步了解植物性化合物與癌症的關聯，本論文第二部份分別以兩株人類乳癌細胞-MCF-7與MDA-MB-231，評估21種台灣蔬果及藥材的抗乳癌能力，並篩選具抗氧化與抗癌活性潛力之天然植物：YC-27，探討其抗兩株乳癌細胞之活性機轉。
< 第二部份 >：分別探討 ( 1 ) 21種台灣蔬果及藥材抗人類乳癌細胞之能力與 ( 2 ) YC-27抗乳癌活性機轉。
( 1 ) 21種台灣蔬果及藥材抗人類乳癌細胞之能力
利用MTT試劑測試天然植物分別對雌激素受體陽性乳癌細胞株MCF-7，與雌激素受體陰性乳癌細胞株MDA-MB-231兩株乳癌細胞之活性機轉。MTT實驗結果顯示在200 μg / ml濃度下，與MCF-7共同培養24小時之抑制細胞生長能力 ( % ) 黑胡椒 = 66.72 ± 5.91和百里香 = 58.95 ± 6.21，以及對MDA-MB-231細胞生長活性能力：YC-27 = 62.16 ± 15.20，皆對乳癌細胞產生細胞活性影響，表現抗乳癌之潛力，並且黑胡椒和百里香 / YC-27於濃度100 μg / ml下，皆使MCF-7 / MDA-MB-231有凋亡小體 ( apoptotic body ) 如細胞萎縮與染色體濃縮及空泡之產生，顯示經由凋亡路徑抑制MCF-7 / MDA-MB-231細胞生長活性。其中YC-27對較惡性之雌激素受體陰性乳癌細胞MDA-MB-231表現抗癌活性，並有良好抗氧化能力，因此進一步探討其抗癌活性機轉。
( 2 ) YC-27之抗乳癌活性及機轉
將YC-27以不同極性溶劑：正己烷 ( Hexane )、乙酸乙酯 ( Ethyl acetate，EA )、丙酮 ( Acetone )、70 % 甲醇 ( 70 % MeOH ) 和甲醇 ( MeOH ) 萃取後，再分別以不同極性溶劑劃分萃取，得到不同極性之濃縮萃取化合物。MTT試驗之細胞活性結果：YC-27萃取物與劃分萃取物於濃度200 μg / ml下，對MCF-7細胞生長抑制率 ( % ) 為：YC-27-E = 76.77 ± 2.94，YC-27-EF1 = 79.29 ± 11.85，YC-27-EF2 = 93.35 ± 3.28，YC-27-A = 54.66 ± 10.54，YC-27-AF1 = 54.47 ± 5.59，YC-27-MF1 = 63.53 ± 6.99，YC-27-MF2 = 91.70 ± 2.59；對MDA-MB-231細胞生長抑制率 ( % ) 為：YC-27-E = 93.80 ± 8.68，YC-27-EF1 = 61.24 ± 5.47，YC-27-A = 97.68 ± 3.18，YC-27-AF1 = 77.40 ± 7.27，YC-27-AF2 = 82.54 ± 4.86，YC-27-M = 62.16 ± 5.20，YC-27-MF2 = 87.08 ± 6.20，YC-27-MF3 = 71.37 ± 3.12，YC-27-70 % M = 65.64 ± 1.40。其中YC-27-AF2、YC-27-M、YC-27-MF3、YC-27-70 % M對兩株乳癌細胞有選擇性抑制能力，具有藥物研發潛力，因此進一步進行細胞活性機轉實驗：以 ( a ) 照相螢光顯微鏡觀察經過染色之細胞型態：YC-27-AF2、YC-27-M、YC-27-MF3、YC-27-70 % M於濃度100 μg / ml 下，皆使MDA-MB-231有細胞萎縮與染色質濃縮及空泡之產生，顯示YC-27經由凋亡路徑抑制MDA-MB-231細胞生長活性。另進一步進行YC-27萃取物與劃分萃取物，與兩株乳癌細胞作用後所產生之 ( b ) 細胞核內去氧核醣核酸斷裂片 ( DNA fragmentation ) 情形： YC-27-MF3產生DNA斷裂 ( c ) 西方墨點轉漬法 ( Western blotting ) 偵測細胞中蛋白質表現變化顯示：YC-27-M、YC-27-MF3與YC-27-70%M於濃度100 μg / ml下，MDA-MB-231皆有Caspase 3蛋白質表現。
綜觀以上結果YC-27具有良好之抗氧化能力，並能抑制兩株不同雌激素受體表現型之乳癌細胞，因此進一步探討YC-27抑制乳癌細胞活性的詳細機轉，並作為抗癌新藥之開發。
Free radicals are correlated with many diseases such as brain and vessel disease、coronary heart disease、diabetes, aging, etc. Polyphenols are the plant-derived compounds which associated with antioxidative and chemopreventive benefits by free-radical reduction. Breast cancer was the top ten death cancer which frequently occurred in past decades. The cases of breast cancer and death rate are still on the rise nowadays. Today, breast cancer is the most common cancer in Taiwan and be associated with women suffering. Therefore, 21 selected Taiwan vegetable,fruit and botany ethanol extractions were investigated their antioxidative potential and anti-cancer proliferative property on two breast cancer cell lines：estrogen-receptor ( ER )-positive breast cancer cell line—MCF-7,and ER-negative breast cancer cell line—MDA-MB-231,to disscuse the diet of breast cancer patient’s in different subtypes. This paper contained the following 2 parts：
< Part I > Free radical scavenging activities and polyphenolic content of selective Taiwanese vegetable,fruit and botany
The polyphenolic contents were evaluated by Total phenol/flavonoid/flavonol quantitative analysis. The results exhibited that the polyphenol-rich plant were： Coffea canephora / C. arabicaJ ( YC-12 )、Thymus vulgaris L. ( YC-13 )、Perilla frutescens L. Britton ( YC-21 )、Mikania cordata Burm. f. B. L. Rob ( YC-24 )、Osmanthus fragrans L. ( YC-25 )、Dimocarpus longan L. ( YC-26 ) and YC-27. And several free-radical-associated assays were used and exhibiting the significant effects with EC50 ( μg / ml ) that show as the following：( 1 ) DPPH free radical scavenging ability：YC-12 = 19.78 ± 0.59、YC-13 = 15.94 ± 1.49、YC-21 = 13.89 ± 1.63、YC-24 = 8.90 ± 2.53、YC-25 = 11.01 ± 0.74、YC-26 = 2.83 ± 0.25 and YC-27 = 19.58 ± 0.36, ( 2 ) Fe2+ chelating ability：Hylocereus costaricensis ( Weber ) Britt. & Rose peel ( YC-14 ) = 75.89 ± 1.93, ( 3 ) XO inhibition：YC-11 = 30.15 ± 6.97, ( 4 ) O2‧- radical scavenging ability：YC-26 = 2.27 ± 0.42 and YC-27 = 33.33 ± 7.59, ( 5 ) LPO inhibition：YC-12 = 47.44 ± 8.09、YC-13 = 7.03 ± 0.44、YC-21 = 98.32 ± 14.24、YC-24 = 81.24 ± 9.30、YC-26 = 12.66 ± 0.70 and YC-27 = 32.91 ± 1.19. And, in concentration of 200 μg / ml,the significant ( 6 ) FRAP ability in trolox equivalent antioxidant capacity ( TEAC ) ( μg / ml ) were：YC-12 = 85.62 ± 1.50、YC-13 = 90.37 ± 1.29、YC-21 = 104.77 ± 1.57、YC-24 = 89.72 ± 1.99、YC-25 = 108.40 ± 2.78 and YC-26 = 98.37 ± 0.42.
< Part II > Human breast cancer cells proliferative activities of selected Taiwanese vegetable, fruit and botany
A. Proliferative activities of selective Taiwanese vegetable,fruit and botany to human breast cancer cell lines—MCF-7 and MDA-MB-231
YC-11 and YC-13 exhibited the inhibitory effect on MCF-7 breast cancer cell proliferation activities with IC50 ( μg / ml ) = 136.67 ± 5.50 and 191.59 ± 0.78. On the other hand, YC-27 inhibited the MDA-MB-231 breast cancer cell proliferation activities with IC50 ( μg / ml ) = 112.79 ± 16.96. Moreover,in cell morphology assay,apoptotic body including cell shrinkage and chromatin condensation were found in 100 μg / ml-YC-11- and 100 μg / ml-YC-13-added MCF-7 cell culture,as well as in 100 μg / ml-YC-27-added MDA-MB-231 cell culture. Liebermann-Burchard test was used for steroid determination and results showed that：steroid-contained samples were YC-11、YC-13、YC-14、YC-20、YC-21、YC-24、YC-25 and YC-26.
B. In vitro anti-tumor genetic effect of YC-27.
Since results of MTT test exhibited YC-27 extraction from methanol ( MeOH ) were significantly inhibit MDA-MB-231 cell line which associated with malignant and metastatic abilities,the botany YC-27 were extracted and fractionated with solvents in different polarity such as hexane,ethyl acetate ( EA ), acetone and MeOH to begin the further exploration. In MTT assay, EA / Acetone / MeOH extraction/fraction at concentration of 200 μg / ml exhibited significantly inhibitory effect ( % ) on ( 1 ) MCF-7 cell：YC-27-E = 76.77 ± 2.94, YC-27-EF1 = 79.29 ± 11.85, YC-27-EF2 = 93.35 ± 3.28 and YC-27-MF2 = 91.70 ± 2.59, and on ( 2 ) MDA-MB-231 cell：YC-27-E = 93.80 ± 8.68, YC-27-EF1 = 61.24 ± 5.47, YC-27-A = 97.68 ± 3.18, YC-27-AF1 = 77.40 ± 7.27, YC-27-AF2 = 82.54 ± 4.86, YC-27-M = 62.16 ± 5.20, YC-27-MF2 = 87.08 ± 6.20, YC-27-MF3 = 71.37 ± 3.12, YC-27-70 % M = 65.64 ± 1.40. Furthermore, in cell morphology assay,apoptotic body including cell shrinkage and chromatin condensation were found in 100 μg / ml-extraction / fraction-added MCF-7 cell culture, as well as in 100 μg / ml-extraction / fraction-added MDA-MB-231 cell culture. In MTT assay, the selected anti-cancer ability between MCF-7 and MDA-MB-231 cell of extractions / fractions－YC-27-AF2、YC-27-M、YC-27-MF3、YC-27-70 % M showed potential of drug development.
DNA fragmentation and western blotting showed that YC-27-MF3 exhibited DNA fragment in DNA fragmentation assay and expresses apoptotic protein－caspase 3 in western blotting assay. In view of antioxidant and anti-breast cancer properties of botany YC-27, the investigation of the potential carcinogenesis and other diseases prevention is feasible and anticipative in the future. |
| 關聯: | 120頁 |
| 描述: | 致謝
目錄．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．1
縮寫表．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．7
表目錄．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．9
圖目錄．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．10
中文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．13
英文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．16
前言．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．19
Part I：台灣蔬果及藥材之自由基清除能力．．．．．．．．．．．．．．． 21
前言．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．22
材料及儀器．．．．．．．．．．．．．．．．．．．．．．．．．．．27
一. 實驗材料．．．．．．．．．．．．．．．．．．．．．．．．27
二. 試藥與試劑．．．．．．．．．．．．．．．．．．．．．．．30
三. 試液之配製．．．．．．．．．．．．．．．．．．．．．．．33
四. 實驗用儀器．．．．．．．．．．．．．．．．．．．．．．．35
實驗方法．．．．．．．．．．．．．．．．．．．．．．．．．．．．37
一. 材料之前處理與萃取．．．．．．．．．．．．．．．．．．．37
二. 總多酚 ( Total Phenolic reagent ) 含量測定．．．．．．．．．．37
三. 總類黃酮 ( Total Flavonoid ) 含量測定．．．．．．．．．．．37
四. 總黃酮 ( Total Flavonol ) 含量測定．．．．．．．．．．．．．37
五. 抗氧化能力測定．．．．．．．．．．．．．．．．．．．．．38
1. 清除DPPH自由基能力測定．．．．．．．．．．．．．．38
2. 螯合亞鐵離子能力測定．．．．．．．．．．．．．．．．38
3. 抑制黃嘌呤氧化酶 ( Xanthine oxidase，XO ) 能力測定．．．39
4. 清除超氧陰離子 ( Superoxide radical，O2‧- ) 能力測定．．．40
5. 清除過氧化氫 ( Hydrogen peroxide，H2O2 ) 能力測定．．．40
6. 還原能力測定 ( Ferric reducing antioxidant power，FRAP )．． 41
六. 脂質過氧化 ( Lipid peroxidation，LPO ) 活性．．．．．．．．．42
實驗結果．．．．．．．．．．．．．．．．．．．．．．．．．．．．44
一. 咖啡、百里香、紫蘇、蔓澤蘭、桂花、龍眼花和YC-27之
總多酚 ( Total Phenol ) 含量大於100 μg Gallic acid / mg．．．．44
二. 咖啡、百里香、紫蘇、蔓澤蘭、龍眼花和YC-27之
總類黃酮 ( Total Flavonoid ) 含量大於10 μg Rutin / mg．．．．45
三. 咖啡、百里香、山藥、紫蘇、蔓澤蘭、龍眼花和YC-27之
總黃酮醇 ( Total Flavonol ) 含量大於20 μg Rutin / mg．．．．．46
四. 21種台灣蔬果及藥材之自由基清除能力結果．．．．．．．．48
1. 咖啡、百里香、原生種山藥、洛神花、紫蘇、蔓澤蘭、
桂花、龍眼花和YC-27有明顯清除DPPH自由基能力．．．．48
2. 咖啡、百里香、紫蘇、蔓澤蘭、桂花和龍眼花有明顯還原能力 ．．．．．．．．．．．．．．．．．．．．．．．．．50
3. 火龍果皮有明顯螯合亞鐵離子能力．．．．．．．．52
4. 黑胡椒有明顯抑制黃嘌呤氧化酶能力．．．．．．．54
5. 苦瓜、洋蔥、百里香、龍眼花和YC-27有明顯
清除超氧陰離子能力．．．．．．．．．．．．．．．．．56
6. 21種天然蔬果及藥材萃取物無明顯清除過氧化氫能力．．．58
五. 21種台灣蔬果及藥材之抑制脂質過氧化 ( LPO ) 活性結果．．．60
1. TEP標準．．．．．．．．．．．．．．．．．．．．．．60
2. 蛋白質定量標準曲線．．．．．．．．．．．．．．．．．60
3. 咖啡、百里香、地瓜、原生種山藥、 山藥、洛神花、
紫蘇、蔓澤蘭、桂花、龍眼花和YC-27有明顯抑制
FeCl2誘導肝粒線體脂質過氧化能力．．．．．．．．．．．61
討論．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．64
Part II：台灣蔬果及藥材對乳癌生長之活性探討．．．．．．．．．．．．．66
前言．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．67
A. 台灣蔬果及藥材對人類乳癌細胞株—MCF-7及MDA-MB-231之生長活性．74
材料及儀器．．．．．．．．．．．．．．．．．．．．．．．．．．．75
一. 實驗材料．．．．．．．．．．．．．．．．．．．．．．．．75
二. 試藥與試劑．．．．．．．．．．．．．．．．．．．．．．．75
三. 試液之配製．．．．．．．．．．．．．．．．．．．．．．．76
四. 實驗用儀器．．．．．．．．．．．．．．．．．．．．．．．77
實驗方法．．．．．．．．．．．．．．．．．．．．．．．．．．．．79
一. 材料之前處理與萃取．．．．．．．．．．．．．．．．．．．79
二. 冷凍細胞之活化．．．．．．．．．．．．．．．．．．．．．79
三. 細胞株之繼代培養．．．．．．．．．．．．．．．．．．．．79
四. 細胞生長活性分析 ( MTT assay )．．．．．．．．．．．．．80
五. 細胞鏡檢反應分析．．．．．．．．．．．．．．．．．．．．81
六. 類固醇 ( Steroid ) 與類三萜 ( Triterpenoid ) 定性測試．．．．．81
實驗結果．．．．．．．．．．．．．．．．．．．．．．．．．．．．82
一. 咖啡在MTT 試驗中表現促進雌激素受體陽性乳癌細胞MCF-7之細胞生長活性，茭白筍莖上葉鞘、火龍果肉和地瓜在
MTT 試驗中表現促進雌激素受體陰性乳癌細胞MDA-MB-231
之細胞生長活性．．．．．．．．．．．．．．．．．．．．．82
二. 黑胡椒和百里香具顯著抑制MCF-7細胞生長活性能力，YC-27
具顯著抑制MDA-MB-231細胞生長活性能力．．．．．．．．82
三. 黑胡椒和百里香使MCF-7產生凋亡小體 ( apoptotic
body ) ．．．．．．．．．．．．．．．．．．．．．．．．85
四. YC-27使MDA-MB-231產生凋亡小體 ( apoptotic
body ) ．．．．．．．．．．．．．．．．．．．．．．．．86
五. 黑胡椒、百里香、火龍果皮、洛神花、紫蘇、蔓澤蘭、桂花
與龍眼花萃取物中含有類固醇 ( Steroid ) 結構之成分．．．．．87
討論．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．88
B. YC-27之抗乳癌細胞活性．．．．．．．．．．．．．．．．．．．．．90
材料及儀器．．．．．．．．．．．．．．．．．．．．．．．．．．．91
一. 實驗材料．．．．．．．．．．．．．．．．．．．．．．．．91
二. 試藥與試劑．．．．．．．．．．．．．．．．．．．．．．．91
三. 試液之配製．．．．．．．．．．．．．．．．．．．．．．．92
四. 實驗用儀器．．．．．．．．．．．．．．．．．．．．．．．93
實驗方法．．．．．．．．．．．．．．．．．．．．．．．．．．．．96
一. 材料之前處理與萃取．．．．．．．．．．．．．．．．．．．96
二. 材料之劃分萃取．．．．．．．．．．．．．．．．．．．96
三. 冷凍細胞之活化．．．．．．．．．．．．．．．．．．．．．96
四. 細胞株之繼代培養．．．．．．．．．．．．．．．．．．．．97
五. 細胞生長活性分析 ( MTT assay ) ．．．．．．．．．．．．．97
六. 細胞鏡檢反應分析．．．．．．．．．．．．．．．．．．．．97
七. 細胞週期分析．．．．．．．．．．．．．．．．．．．．．．97
八. 西方點墨法 ( Western blotting ) ．．．．．．．．．．．．．．97
九. 核酸電泳法 ( DNA fragmentation ) ．．．．．．．．．．．．．98
十. 類固醇 ( Steroid ) 與類三萜 ( Triterpenoid ) 定性測試．．．．．99
實驗結果．．．．．．．．．．．．．．．．．．．．．．．．．．．．100
一、 YC-27-MF2、YC-27-E、YC-27-EF1和YC-27-EF2具顯著
抑制MCF-7細胞生長活性能力，YC-27-MF2、YC-27- MF3、
YC-27-E、YC-27-AF1和YC-27-AF2具顯著抑制MDA-MB-231
細胞生長活性能力．．．．．．．．．．．．．．．．．．．．100
二、 YC-27-E、YC-27-EF1 、YC-27-EF2、YC-27-A 、YC-27-AF1 、
YC-27-MF1 和YC-27-MF2使MCF-7產生凋亡小體
( apoptotic body ) ．．．．．．．．．．．．．．．．．．．．102
三、 YC-27-E、YC-27-EF1、YC-27-A、YC-27-AF1、YC-27-AF2、
YC-27-M、YC-27-MF2 、YC-27-MF3和YC-27-70%M
使MDA-MB-231產生凋亡小體 ( apoptotic body ) ．．．．．．102
四、 YC-27-E、YC-27-EF1 、YC-27-EF2、YC-27-A 、YC-27-AF1 、
YC-27-MF1 和YC-27-MF2於MCF-7細胞表現Sub-G1．．．．．105
五、 YC-27-E、YC-27-EF1、YC-27-A、YC-27-AF1、YC-27-AF2、
YC-27-M、YC-27-MF2 、YC-27-MF3和YC-27-70 % M
於MDA-MB-231細胞表現Sub-G1．．．．．．．．．．．．．105
六、 YC-27-H、YC-27-EF1、YC-27-A、YC-27-AF1、
YC-27-MF1、YC-27-MF2與YC-27-70%M萃取物中含有植物性
類固醇 ( Steroid ) 結構之成分．．．．．．．．．．．．．．．108
七、 YC-27-MF3、YC-27-M與YC-27-70%M 能誘導MDA-MB-231
表現Caspase 3蛋白質．．．．．．．．．．．．．．．．．．109
八、 YC-27-MF3使MDA-MB-231產生DNA斷裂片段．．．．．．．110
討論．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．113
參考文獻．．．．．．．．．．．．．．．．．．．．．．．．．．．．．．115
1. Azizova OA, Sergienko VI, Syrkin AL, Ivanov GG, Aseĭchev AV, Lopukhin IuM. ( 2009 ) Clinical and prognostic significance of free radical processes in patients with coronary heart disease. Vestn Ross Akad Med Nauk. ( 10 ):32-40.
2. Błasiak J, Skłodowska A, Ulińska M, Szaflik JP. ( 2009 ) Iron and age-related macular degeneration. Klin Oczna. 111 ( 4-6 ):174-7.
3. Barnes S. ( 2010 ) The biochemistry, chemistry and physiology of the isoflavones in soybeans and their food products. Lymphat Res Biol. 8 ( 1 ):89-98.
4. Beckman KB, Ames BN. ( 1997 ) Oxidative decay of DNA. J Biol Chem. 272 ( 32 ):19633-6
5. Birnboim HC. ( 1988 ) A superoxide anion induced DNA strand-break metabolic pathway in human leukocytes:effects of vanadate. Biochem Cell Biol. 66 ( 5 ):374-81.
6. Bravo L. ( 1998 ) Polyphenols: chemistry, dietary sources, metabolism, and nutritional significance. Nutr Rev. 56:317-33
7. Brisdelli F, D'Andrea G, Bozzi A. ( 2009 ) Resveratrol: a natural polyphenol with multiple chemopreventive properties. Curr Drug Metab. 10 ( 6 ):530-46.
8. Cao G, Sofic E, Prior RL. ( 1997 ) Antioxidant and prooxidant behavior of flavonoids: structure-activity relationships. Free Radic Biol Med. 22 ( 5 ):749-60.
9. Chueh PJ, Wu LY, Morré DM and Morré DJ. ( 2004 ) tNOX is both necessary and sufficient as a cellular target for the anticancer actions of capsaicin and the green tea catechin (-)-epigallocatechin-3-gallate. Biofactors. 20 ( 4 ):235-49.
10. Dapkevicius A, van Beek TA, Lelyveld GP, van Veldhuizen A, de Groot A, Linssen JP, Venskutonis R. ( 2002 ) Isolation and structure elucidation of radical scavengers from Thymus vulgaris leaves. J Nat Prod. 65 ( 6 ):892-6.
11. Deng G, Davatgarzadeh A, Yeung S, Cassileth B. ( 2010 ) Phytoestrogens: science, evidence, and advice for breast cancer patients. J Soc Integr Oncol. 8 ( 1 ):20-30.
12. Ellinger S, Ellinger J, Müller SC, Stehle P. ( 2009 ) Tomatoes and lycopene in prevention and therapy--is there an evidence for prostate diseases? Aktuelle Urol. 40 ( 1 ):37-43.
13. Feskens EJ, and Katan MB. ( 1999 ) Tea flavonols in cardiovascular disease and cancer epidemiology. Proc Soc Exp Biol Med 220,198-202
14. Floyd RA. ( 1999 ) Antioxidants, oxidative stress, and degenerative neurological disorders. Proc Soc Exp Biol Med.222:236-245
15. Fröjdö S, Durand C, Pirola L. ( 2008 ) Metabolic effects of resveratrol in mammals--a link between improved insulin action and aging. Curr Aging Sci. 1 ( 3 ):145-51.
16. Fuster JJ, Andrés V. ( 2006 ) Telomere biology and cardiovascular disease. Circ Res. 99 ( 11 ):1167-80.
17. Gamal-Eldeen AM, Djemgou PC, Tchuendem M, Ngadjui BT, Tane P, Toshifumi H. ( 2007 ) Anti-cancer and immunostimulatory activity of chromones and other constituents from Cassia petersiana. Z Naturforsch C. 62 ( 5-6 ):331-8.
18. Hakim FA, Pflueger A. ( 2010 ) Role of oxidative stress in diabetic kidney disease. Med Sci Monit. 16 ( 2 ):RA37-48.
19. Harrison DG, Gongora MC. ( 2009 ) Oxidative stress and hypertension. Med Clin North Am. 93 ( 3 ):621-35.
20. Higashi Y. ( 2009 ) Edaravone for the treatment of acute cerebral infarction: role of endothelium-derived nitric oxide and oxidative stress. Expert Opin Pharmacother. 10 ( 2 ):323-31.
21. Ho HY, Cheng ML, Weng SF, Leu YL, Chiu DT. ( 2009 ) Antiviral effect of epigallocatechin gallate on enterovirus 71. J Agric Food Chem. 57 ( 14 ):6140-7
22. Hollman PC, Hertlog MG, Kalan MB. ( 1996 ) Role of dietary flavinoids in protection against cancer and coronary heart disease. Biochem Soc Trans. 24: 785-89.
23. Hsieh TC, Wu JM. ( 2009 ) Targeting CWR22Rv1 prostate cancer cell proliferation and gene expression by combinations of the phytochemicals EGCG, genistein and quercetin. Anticancer Res. 29 ( 10 ):4025-32.
24. I Singh R, Singh N, Saini BS, Rao HS. ( 2008 ) In vitro antioxidant activity of pet ether extract of black pepper. Indian J Pharmacol. 40 ( 4 ):147-51.
25. Kawada H, Kojima M, Kimura T, Natori S, Sasaki K, Sasaki H. ( 2009 ) Effect of 5-S-GAD on UV-B-induced cataracts in rats. Jpn J Ophthalmol. 53 ( 5 ):531-5.
26. Kidd PM. ( 2009 ) Bioavailability and activity of phytosome complexes from botanical polyphenols: the silymarin, curcumin, green tea, and grape seed extracts. Altern Med Rev. 14 ( 3 ):226-46.
27. Kotan R, Cakir A, Dadasoglu F, Aydin T, Cakmakci R, Ozer H, Kordali S, Mete E, Dikbas N. ( 2010 ) Antibacterial activities of essential oils and extracts of Turkish Achillea, Satureja and Thymus species against plant pathogenic bacteria. J Sci Food Agric. 90 ( 1 ):145-60.
28. Kovacic P, Pozos RS, Somanathan R, Shangari N, O'Brien PJ. ( 2005 ) Mechanism of mitochondrial uncouplers, inhibitors, and toxins: Focus on electron transfer, free radicals, and structure-activity relationships. Curr Med Chem. 12 ( 22 ):2601-23.
29. Kovacic P, Jacintho JD. ( 2001 ) Mechanisms of carcinogenesis:focus on oxidative stress and electron transfer. Curr Med Chem. 8, pp.773-96.
30. Lamoral-Theys D, Pottier L, Dufrasne F, Nève J, Dubois J, Kornienko A, Kiss R, Ingrassia L. ( 2010 ) Natural polyphenols that display anticancer properties through inhibition of kinase activity. Curr Med Chem. 17 ( 9 ):812-25.
31. Lee DW, Andersen JK. ( 2010 ) Iron elevations in the aging Parkinsonian brain: a consequence of impaired iron homeostasis? J Neurochem. 112 ( 2 ):332-9.
32. Lelono RA, Tachibana S, Itoh K. ( 2009 ) In vitro antioxidative activities and polyphenol content of Eugenia polyantha Wight grown in Indonesia. Pak J Biol Sci. 15;12 ( 24 ):1564-70.
33. Li N, Zhao B, Yu YF, Dong XP. ( 2008 ) Studies on anti-inflammation chemical constitutes of Patrinia villosa. Zhong Yao Cai. 31 ( 1 ):51-3.
34. Valko M, Izakovic M, Mazur M, Rhodes CJ, Telser J. ( 2004 ) Role of oxygen radicals in DNA damage and cancer incidence. Mol. Cell. Biochem. 266 ( 1-2 ), 37-56.
35. Manuel JS and Maria del Carmen MS. ( 2009 ) Suppression of Met activation in human colon cancer cells treated with (-)-epigallocatechin-3-gallate: minor role of hydrogen peroxide. Biochem Biophys Res Commun. 389 ( 3 ):527-30.
36. Jimenez-Saenz M, Martinez-Sanchez Mdel C. ( 2006 ) Acute hepatitis associated with the use of green tea infusions. J Hepatol 44;616–17
37. Martí N, Mena P, Cánovas JA, Micol V, Saura D. ( 2009 ) Vitamin C and the role of citrus juices as functional food. Nat Prod Commun. 4 ( 5 ):677-700.
38. Mecocci P, Fanó G, Fulle S, MacGarvey U, Shinobu L, Polidori MC, Cherubini A, Vecchiet J, Senin U, Beal MF. ( 1999 ) Age-dependent increases in oxidative damage to DNA,lipids,and proteins in human skeletal muscle. Free Radic Biol Med. 26:303-308.
39. Morré DM, Morré DJ. ( 2006 ) Anticancer activity of grape and grape skin extracts alone and combined with green tea infusions. Cancer Lett. 18;238 ( 2 ):202-9.
40. Namiki M. ( 2007 ) Nutraceutical functions of sesame: a review. Crit Rev Food Sci Nutr. 47 ( 7 ):651-73.
41. Pace-Asciak CR, Hahn S, Diamandis EP, Soleas G, Goldberg DM. ( 1995 ) Wines and grape juices as modulators of platelet aggregation in healthy human subjects. Clin Chim Acta. 235:207-19
42. Pannala AS, Rice-Evans C. ( 2001 ) Rapid screening method for relative antioxidant activities of flavonoids and phenolics. Methods Enzymol. 335:266-72.
43. Peterson J and Dwyer J. ( 1998 ) Flavonoids: dietary occurrence and biochemical activity. Nutr. Res. 18:1995-2018.
44. Rice-Evans CA and NJ Miller. ( 1996 ) Antioxidant activities of flavonoids as bioactive components of food. Biochem. Soc. Trans. 24: 790-95.
45. Ridnour LA, Thomas DD, Mancardi D, Espey MG, Miranda KM, Paolocci N, Feelisch M, Fukuto J, Wink DA. ( 2004 ) The chemistry of nitrosative stress induced by nitric oxide and reactive nitrogen oxide species. Putting perspective on stressful biological situations. Biol Chem. 385 ( 1 ):1-10.
46. Sanbongi C, Suzuki N and Sakane T. ( 1997 ) Polyphenols in chocolate, which have anti-oxidant activity, modulate immune functions in humans in vitro. Cell. Immunol. 177:129-36.
47. Suzuki M, Tabuchi M, Ikeda M, Umegaki K, Tomita T. ( 2004 ) Protective effects of green tea catechins on cerebral ischemic damage. Med Sci Monit. 10 ( 6 ):BR166-74
48. Tada Y, Suga K, Imai K, Nakachi K. ( 1998 ) Cancer Inhibition by Green Tea. Mutation Research 402:307-310.
49. Vasanthi P, Nalini G, Rajasekhar G. ( 2009 ) Status of oxidative stress in rheumatoid arthritis. Int J Rheum Dis. 12 ( 1 ):29-33.
50. Wan Y, Vinson JA, Etherton TD, Proch J, Lazarus SA, Kris-Etherton PM. ( 2001 ) Effects of cocoa powder and dark chocolate on LDL oxidative susceptibility and prostaglandin concentrations in humans. Am J Clin Nutr 74:596-602.
51. Waterhouse AL, Shirley JB, Donovan JL. ( 1996 ) Antioxidants in chocolate. Lancet. 348:834.
52. Yang CS, Chung JY, Yang G, Chhabra SK and Lee M-J. ( 2000 ) Tea and tea polyphenols in cancer prevention. J. Nutr. 130:472S-78S
53. Yi-Zhong Cai, Mei Sun, Jie Xing, Qiong Luo and Harold Corke. ( 2006 ) Structure-radical scavenging activity relationships of phenolic compounds from traditional Chinese medicinal plants. Life sciences 78 ( 25 ):2872-88
54. Zuercher AW, Holvoet S, Weiss M, Mercenier A. ( 2010 ) Polyphenol-enriched apple extract attenuates food allergy in mice. Clin Exp Allergy. 40 ( 6 ):942-50. |
| 顯示於類別: | [生藥學研究所] 博碩士論文
|
在TMUIR中所有的資料項目都受到原著作權保護.
|
