台灣多酚植物(YA-1)之抗氧化及肝損傷保護作用

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题名:	台灣多酚植物(YA-1)之抗氧化及肝損傷保護作用
作者:	王啟任
贡献者:	生藥學研究所
日期:	2010
上传时间:	2010-10-21 09:10:58 (UTC+8)
摘要:	現代社會的複雜經濟結構，人們每天只顧忙碌地工作，一般人對健康較為疏忽，因此肝病ㄧ直是台灣國人健康的殺手，根據衛生署統計，在台灣每年約有五千人死於肝癌，四千人死於肝硬化和慢性肝炎，甚至於從民國七十年以來，肝癌一直是國人男性癌症死因的首位，女性癌症死因的第二位。由於肝臟沒有神經，多數的肝病病人不會感到疼痛，也沒有其他症狀，當病人感到身體異常而就醫時，往往已是肝癌末期，為此，對於肝臟的保護與肝病的預防更顯得重要，正常的生活作息與均衡飲食能有效預防肝臟疾病。目前臨床常見治療肝臟疾病與肝癌的方法，包含了：手術切除、經動脈栓塞術療法、經皮酒精注射療法、經內視鏡食道靜脈瘤硬化療法、經內視鏡靜脈瘤結扎術、肝移植與藥物治療，但停藥後復發率極高。根據研究指出，合併服用抗氧化劑能提升肝臟復原效果，降低藥物的副作用與停藥後的復發率。現今科學證實植物多酚具有抗氧化能力，台灣擁有豐富的植物資源，善加利用台灣之優勢亦為本研究之目標。 YA-1為落葉喬木，樹幹粗大，質地堅硬，在台灣僅有2種，主要的差異在於葉子的大小，分別是YA-1S(小葉)和YA-1L(大葉)，小葉採自台南，大葉採自雲林、南投與屏東，此篇論文以五種不同極性的溶劑進行萃取，包含：正己烷(HE)、乙酸乙酯(EE)、丙酮(AE)、甲醇(ME)，70%甲醇(70ME)。利用抗氧化與抑制氯化亞鐵誘導肝臟粒線體脂質過氧化進行體外護肝能力評估，其中以雲林YA-1L-ME有最佳之抑制率，再以四種不同溶劑萃取，分別是：正己烷(HF)、乙酸乙酯(EF)、丙酮(AF)、甲醇(MF)。本實驗評估總多酚、總黃酮與黃烷醇含量，以及抗氧化能力、抑制肝臟粒線體脂質過氧化能力，其中以雲林YA-1L-AF萃取物效果最好。再分別選出有效抑制肝臟粒線體脂質過氧化之萃取物，包含：台南YA-1S-AE、台南YA-1S-ME、雲林YA-1L-ME與雲林YA-1L-AF，接著利用四氯化碳誘導ICR小鼠急性肝損傷評估萃取物在活體內保護肝臟之能力。從血清生化檢測中顯示雲林YA-1L-AF能有效降低血清中GOT、GPT值，另由病理組織切片分析發現亦能減少肝細胞壞死現象，同時，能有效保護肝臟粒線體DNA之受損。因此，雲林YA-1L具有潛力成為護肝保健食品或新藥物之開發。
關聯:	106頁
描述:	致謝………………………………………………………………………1 目錄……………………………………………………………………2 表目錄…………………………………………………………………6 圖目錄…………………………………………………………………8 縮寫表…………………………………………………………………10 中文摘要………………………………………………………………12 英文摘要………………………………………………………………14 前言……………………………………………………………………16 試劑與儀器……………………………………………………………32 一、試藥與試劑…………………………………………………32 二、實驗儀器……………………………………………………35 三、試劑配製……………………………………………………37 四、植物材料……………………………………………………40 五、實驗動物……………………………………………………40 實驗方法………………………………………………………………41 一、植物材料前處理與萃取…………………………………41 二、氯化亞鐵(FeCl2)誘導大鼠肝臟粒腺體酯質過氧化之抑制能力…………………………………………………………42 (1)蛋白質定量……………………………………………42 (2)TEP標準曲線…………………………………………43 (3)YA-1於肝臟粒腺體酯質過氧化活性測定……………43 (4)抑制酯質過氧化計算公式……………………………44 三、四氯化碳(CCl4)誘導ICR小鼠急性肝損傷………………44 (1)YA-1萃取物對小鼠急性肝損傷之血清生化檢測……44 (2)YA-1萃取物對小鼠急性肝損傷之病理組織切片……45 (3)YA-1萃取物對肝臟粒線體DNA之保護……………45 四、總多酚、總黃酮與總黃烷醇含量測定……………………47 (1)總多酚含量測定(Total phenol)…………………………47 (2)總黃酮含量測定(Total flavonoid)………………………48 (3)總黃酮醇含量測定(Total flavonol)……………………48 五、自由基清除與抗氧化活性評估……………………………49 (1)清除DPPH自由基能力測定……………………………49 (2)螯合亞鐵離子能力測驗………………………………50 (3)清除超氧陰離子能力測定……………………………50 (4)抑制超氧陰離子能力測定……………………………51 (5)總還原能力測定………………………………………52 六、統計分析……………………………………………………53 實驗結果………………………………………………………………54 一、氯化亞鐵(FeCl2)誘導大鼠肝臟粒腺體酯質過氧化之抑制能力……………………………………………………………54 (1) MDA(TBA)2含量標準品TEP之標準曲線…………54 (2) 蛋白質定量標準品BSA之標準曲線………………55 (3) YA-1抑制肝臟粒線體脂質過氧化能力……………55 二、四氯化碳(CCl4)誘導ICR小鼠急性肝損傷之血清生化檢測……………………………………………………………59 三、四氯化碳(CCl4)誘導ICR小鼠急性肝損傷之病理組織切片……………………………………………………………61 四、YA-1萃取物對肝臟粒線體DNA之保護…………………65 五、總多酚含量…………………………………………………67 六、總黃酮含量…………………………………………………69 七、總黃酮醇含量………………………………………………71 八、清除DPPH自由基能力……………………………………72 九、螯合亞鐵離子能力…………………………………………75 十、清除超氧陰離子能力………………………………………78 十一、抑制超氧陰離子能力……………………………………81 十二、總還原能力………………………………………………83 討論……………………………………………………………………85 結論……………………………………………………………………89 參考文獻………………………………………………………………91 附錄…………………………………………………………..………100 Aaron J. 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