臺灣博碩士論文加值系統

全球每年約有 65.5 萬人死於結直腸癌。流行病學研究顯示，飲食中攝取含有大量類黃酮物質的水果與蔬菜，可能有助於預防人類結直腸癌的發生。根據研究，海巴戟天葉乙醇萃取物（LES）及其分劃物（F I-V）、香蓼嫩枝甲醇 （PoM） 或水（PoW）萃取物，除了富含高多酚類化合物之外，更擁有良好抗氧化活性。因此，本研究進一步探討LES、F I-V、PoM 及 PoW 對結直腸癌細胞(HT-29) 的抗癌活性，是否可藉由活化caspase-3，以及調節細胞核轉錄因子NF-kappa B 或 AP-1 的表現而達成。研究結果顯示 LES、F I、F III (Rutin) PoM 及 PoW 皆可抑制 HT-29的細胞增生，以及增加與凋亡機制相關的蛋白質 AP-1 及 caspase-3 的表現。此外，F I 在細胞存活率及基因毒性試驗中，誘導 HT-29 細胞凋亡及細胞週期之影響的結果與 LES 結果類似；然而，PoW 在抑制HT-29 細胞的增生、NF-kappa B 的表現以及 caspase-3 的活化較優於 PoM 。因此，F I 與 PoW 可為抗癌活性的重要來源，值得進一步純化與鑑定其生物活性成分，並深入探討其抗癌機制。此外，清楚了解 F I 與 PoW 的生物活性機制，有助於發展成為保健食品或治療藥物。

With 655,000 deaths in worldwide is due to colorectal carcinoma per year. Epidemiological studies suggest that ingesting a lot of dietary flavonoids with fruits and vegetables may prevent the incidene of colorectal carcinoma in humans. Based on our researches, ethanolic extracts from Morinda citrifolia leaf (LES), methanolic (PoM) or aqueous (PoW) extracts from Polygonum odoratum young shoots (Po) exhibited higher polyphenolic contents and revealed that these compounds played a part in antioxidative activities. LES, sub-fractions from LES (F I-V), PoM and PoW were used to study anti-cancer activity by acting through caspase-3 and modulating the expressions of NF-kappa B or AP-1 in colorectal carcinoma cells (HT-29), respectively. Results suggested that LES, F I, F III (rutin), PoM and PoW inhibited the proliferation of HT-29 and was associated with an apoptotic mechanism involving increasing in AP-1 expression and caspase-3 activation. Moreover, effects of F I on cell viability, genotoxicity, induction of apoptosis and cell cycle arrest were all similar to those of LES in HT-29. However, inhibition of HT-29 proliferation, and NF-kappa B expression as well as caspase-3 activation of PoW were better than PoM. Therefore, F I and PoW should be significant sources of anti-cancer activity. It was worth further research in purification and identification of active compounds, and deeply investigation for anti-cancer effects. Moreover, a clear understanding of the molecular mechanisms of action of F I and PoW are crucial in the valuation of these potent molecules as potential prophylactic and therapeutic agents.

中文摘要 I
英文摘要 II
目錄 IV
表目錄 VII
圖目錄 VIII
縮寫表 X
第一章 緒論 1
第一節 結直腸癌 1
一、結直腸癌的盛行率 1
二、結直腸的位置與生理功能 2
三、結直腸癌的好發區域 2
四、結直腸癌的高危險群及危險因子 3
五、結直腸癌的治療 4
第二節 多酚類化合物對癌症的影響 4
第三節 細胞的生長與死亡 11
一、 細胞週期的調控 11
二、凋亡在人類疾病中扮演的角色 11
三、 細胞凋亡調控路徑 12
四、 細胞內轉錄因子調控細胞凋亡 14
第五節 本研究所採用之天然萃取物 17
一、海巴戟天 17
二、 香蓼 28
三、 海巴戟天葉及香蓼之綜合討論 29
第六節 研究動機與目的 31
第二章 研究設計與材料方法 32
第一節 實驗設計與架構 32
第二節 材料製備 33
一、 海巴戟天葉乙醇萃取物 LES 及其分劃物之製備 33
二、 香蓼嫩枝水萃 (PoW) 及甲醇萃取物 (PoM) 之製備 35
第三節 實驗分析項目及其測定方法 36
一、細胞培養 36
二、細胞存活率試驗 37
三、總抗氧化能力測定 38
四、凋亡相關蛋白檢測 38
五、細胞內轉錄因子分析 40
六、基因毒性分析 48
七、細胞凋亡與細胞壞死測定 52
八、檢測細胞週期的分布情形 54
九、數據處理 56
第三章 結果 57
第一節 測試樣品對 HT-29 細胞存活率影響 57
第二節 測試樣品之總抗氧化能力檢測 59
第三節 測試樣品誘導 HT-29 細胞產生 ROS 59
第四節 測試樣品對 HT-29 細胞之基因毒性 60
第五節 測試樣品對凋亡相關蛋白的影響 61
第六節 測試樣品對細胞轉錄因子 NF-?羠 影響 63
第七節 測試樣品對細胞轉錄因子 AP-1 影響 64
第八節 測試樣品誘發 HT-29 細胞凋亡的效應 65
第九節 測試樣品對 HT-29 細胞之細胞週期之影響 66
第四章 討論 69
第五章 結論 78
參考文獻 79
附圖與附表 91

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