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研究生:黃靖容
研究生(外文):Chin Jung Huang
論文名稱:香椿之抗氧化及抗癌活性之探討
論文名稱(外文):Antioxidant and anticancer activities of aqueous extracts from Toona sinensis
指導教授:楊新玲 許游章
指導教授(外文):Hsin Ling Yang, You Cheng Hseu
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱: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:2005
畢業學年度:93
語文別:中文
論文頁數:215
中文關鍵詞:香椿抗氧化劑低密度脂蛋白動脈粥狀硬化人類急性骨髓血癌細胞細胞凋亡
外文關鍵詞:Toona sinensisAntioxidantLow density lipoproteinAtherosclerosisHuman premyelocytic leukemia cellsApoptosis
相關次數:
  • 被引用被引用:20
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中文總摘要
  香椿(Toona sinensis Roem.)為楝科(Meliaceae)之落葉喬木。香椿的嫩葉可食,是自古以來深受喜愛的本土蔬菜,根據文獻資料可知全樹各部分均有保健或治療之功能。研究發現香椿中含有酚類及類黃酮化合物,並具有抗癌及降低血糖之效用。本研究中,將以香椿為主要題材,分別探討其抗氧化活性,抑制低密度脂蛋白氧化及抗癌作用三部分。
  首先以香椿萃取液進行體外的抗氧化活性分析。結果顯示香椿萃取液在亞麻油酸乳化系統中具有很強的抗氧化性及很好的還原力、清除DPPH 自由基、清除超氧陰離子及螯合亞鐵離子的能力。而且,抗氧化能力隨著香椿萃取液濃度增加而增加。由結果可知,香椿萃取液具有良好的抗氧化能力,可能具有開發預防自由基相關疾病之抗氧化劑潛力。
  第二部分探討香椿萃取液保護自由基誘發劑(CuSO4、AAPH、SNP) 誘導低密度脂蛋白(Low density lipoprotein;LDL)過氧化作用。結果顯示香椿萃取液可抑制低密度脂蛋白之氧化作用,包括降低脂質過氧化產物-丙二醛(Malondialdehyde;MDA)生成量及防止膽固醇氧化作用,同時也減少LDL受到氧化修飾所造成的Apoprotein B蛋白裂解現象及LDL蛋白表面電荷改變的程度。這顯示出香椿萃取液可抑制低密度脂蛋白之氧化作用,可能可以預防心血管疾病之發生。
  第三在抗癌部分,以香椿萃取液作用於人類急性骨髓血癌細胞(Human Premyelocytic Leukemia Cells;HL-60),探討香椿萃取液抑制HL-60細胞存活率、生長作用及其可能機轉,並檢測其是否對人類正常內皮細胞具細胞毒殺性。結果發現,香椿萃取液在對正常內皮細胞無傷害的情況下,會減少HL-60細胞存活率及生長情形,且在顯微鏡的觀察下, 細胞有減少和皺縮現象,同時藉由西方墨點法(Western blotting)觀察以香椿萃取液處理之HL-60細胞,其蛋白質表現量的變化,結果顯示Bcl-2表現量減少,Bax、cytochrome c蛋白質量增加,Caspase 3被活化且PARP被分解成兩片段(116和89 kDa)。由以上結果顯示香椿萃取液對HL-60細胞具有藉由誘導細胞凋亡而表現出有效的抗增殖性,應該具有開發成人類血癌治療藥物的潛力。
  總結,香椿萃取液具有良好的抗氧化能力,且可保護低密度脂蛋白之氧化作用並誘導人類急性骨髓血癌細胞(HL-60)細胞凋亡之功效。
英文總摘要
Toona sinensis Roem belongs to the family Meliacceae.The leaves and young shoots are used as vegetable in Chin.In fact,the trees are also widely used medically.Previous phytochemical work on Toona sinensis had led to the isolation phenolic compounds. To further search for the novel bioactive agents from Meliaceae plants, Toona sinensis was chosen for phytochemical investigation and known compounds including, gallic acid, methyl gallate, kaempferol, quercitin, quercitrin, rutin and catechin, were isolated and identified from this plant.
In this study, we have investigated the antioxidant properties of the aqueous extracts of whole fresh Toona sinensis were evaluated using different antioxidant tests, including reducing power ,free radical scavenging, superoxide anion radical scavenging, and metal chelating activities. The results showed that the aqueous extracts of whole fresh Toona sinensis had antioxidative activites in linoleic acid peroxidation system.Moreover, the aqueous extracts of whole fresh Toona sinensis had effective reducing power, free radical scavenging, superoxide anion radical scavenging, and metal chelating activities at the same concentrations.
We have also investigated the effects (antioxidant properties) of Toona sinensis on the oxidative modification of human low-density lipoproteins (LDL), as induced by copper sulfate (CuSO4), 2,2-azobis-2-amidinopropane hydrochloride (AAPH), and sodium nitroprusside (SNP). Under such oxidant stress, Toona sinensis appear to possess antioxidant properties with respect to oxidation of LDL in a time- and concentration-dependent manner, as assessed by inhibition of thiobarbituric acid-reactive substances (TBARS) formation and cholesterol degradation of oxidized LDL. In addition , Toona sinensis exhibited a remarkable ability to rescue the relative electrophoretic mobility and fragmentation of the Apo B moiety of the oxidized LDL. Our findings suggest that the antioxidant properties of Toona sinensis may also provide effective protection from atherosclerosis.
In this study, the ability of Toona sinensis to induce apoptosis was studied in cultured human premyelocytic leukemia HL-60 cells. Treatment of the HL-60 cells with a variety of concentrations of aqueous crude extracts of Toona sinensis (10-75 mg/ml) resulted in dose- and time-dependent sequences of events marked by apoptosis, as shown by loss of cell growth and viability,cell shrinkage. Furthermore, apoptosis in the HL-60 cells was accompanied by the release of cytochrome c, activation of caspase 3 and specific proteolytic cleavage of poly (ADP-ribose) polymerase (PARP). This increase in Toona sinensis induced apoptosis was also associated with a reduction in the levels of Bcl-2, a potent cell-death inhibitor, and an increase in those of the Bax protein, which heterodimerizes with and thereby inhibits Bcl-2. The data suggest that Toona sinensis exerts antiproliferative action and growth inhibition on HL-60 cells through apoptosis induction, and that it may have anticancer properties valuable for application in drug products.
中文總摘要
英文總摘要
論文研究架構圖
第一部分香椿萃取液抗氧化活性之探討
第一章、中文摘要……………………………………………………………………….3
第二章、文獻整理及研究動機………………………………………………………….4
第三章、實驗設計架構圖………………………………………………………………34
第四章、實驗器材………………………………………………………………………36
第五章、實驗方法………………………………..........................38
第六章、實驗結果………………………………………………………………………45
第七章、討論及結論……………………………………………………………………48
第八章、圖表……………………………………………………………………………53
第九章、參考文獻………………………………………………………………………60
第二部分香椿萃取液抑制低密度脂蛋白過氧化之探討
第一章、中文摘要………………………………………………………………………78
第二章、文獻整理及研究動機………………………………………………………….79
第三章、實驗設計架構圖……………………………………………………………..101
第四章、實驗器材……………………………………………………………………..103
第五章、實驗方法……………………………… ..........................106
第六章、實驗結果……………………………………………………………………..114
第七章、討論及結論..…………………………………………………………………118
第八章、圖表…………………………………………………………………………..122
第九章、參考文獻……………………………………………………………………..132
第三部分香椿萃取液誘導人類急性骨髓血癌細胞(Human Premyelocytic Leukemia Cells;HL-60)細胞凋亡之探討
第一章、中文摘要………………………………………………………………………145
第二章、文獻整理及研究動機…………………………………………………………147
第三章、實驗設計架構圖………………………………………………………………166
第四章、實驗器材………………………………………………………………………168
第五章、實驗方法………………………………..........................171
第六章、實驗結果………………………………………………………………………183
第七章、討論及結論……………………………………………………………………188
第八章、圖表……………………………………………………………………………194
第九章、參考文獻………………………………………………………………………201
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