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研究生:廖東垣
研究生(外文):Liao Tung-Yuan
論文名稱:巴蔘免疫調節與抗氧化活性之研究
論文名稱(外文):Studies on the Immunomodulatory and Antioxidant Activities of Talinum fruticosum(L.) Juss
指導教授:蔡明勳蔡明勳引用關係
指導教授(外文):Tsai Ming-Hsun
口試委員:林麗雲林耀東張基郁顏裕鴻蔡明勳
口試委員(外文):Lin Li-YunLin Yao-TungChang Chi-YuYen Yu-HongTsai Ming-Hsun
口試日期:2013-09-18
學位類別:博士
校院名稱:大葉大學
系所名稱:生物產業科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:102
中文關鍵詞:巴蔘萃取物抗氧化活性人類單核球細胞人類白血病細胞U937免疫調節活性
外文關鍵詞:Talinum fruticosum extractsAntioxidant activitiesHuman mononuclear cellsHuman leukemic U937 cellsImmuno-modulatory activity
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本研究以不同溶劑,包括磷酸緩衝溶液、75%乙醇、90℃的沸水,進行巴蔘莖與葉之萃取,以探討萃取物的抗氧化功效,及對人類單核球細胞之增生、誘導細胞激素(IL-1β、IFN-γ及TNF-α)和一氧化氮的分泌和對人類白血病細胞U937生長抑制之影響。結果顯示巴蔘莖與葉的三種溶劑萃取物除了具有顯著的抗氧化活性,利用90℃的熱水萃取所得之巴蔘莖萃取物(WTS)最能顯著刺激人類單核球細胞之增生,在刺激細胞激素分泌方面,以濃度為500 μg/mL的WTS處理人類單核球細胞3天後所得之條件培養液中,其IL-1β、TNF-α及IFN-γ之含量分別為1275、2222和859 pg/mL。所有巴蔘萃取物處理所得之人類單核球細胞培養液皆能顯著抑制人類白血病U937細胞的生長,其抑制率皆為40%以上。根據上述之結果,巴蔘萃取物可應用於免疫調節健康食品之開發。

In this study, leaves and stems of Talinum fruticosum were sequentially extracted with phosphate buffer solution (PTL and PTS), 75% ethanol(ETL and ETS), and 90℃ boiling water(WTL and WTS), respectively. We investigated antioxidant activities of various Talinum fruticosum extracts, analyzed the extracts’ stimulations on human mononuclear cell (MNC) growth and secretion of cytokines (IL-1β, IFN-γ and TNF-α) and nitric oxide, and then assayed their subsequent inhibitions to human leukemic U937 cell growth. Results indicated that obtained extracts of Talinum fruticosum showed significant antioxidant activities. Among the extracts, WTS showed the most significant stimulation on human MNC growth. IL-1β, IFN-γ and TNF-α secretion levels in a conditioned medium that human MNC was treated with 500 μg/mL WTS for 72 hours were 1275, 859 and 2222 pg/mL respectively. All conditioned medium that obtained from human MNC cultured with various Talinum fruticosum extracts, showed significant inhibition against U937 cell growth over 40%. These results suggest that Talinum fruticosum extracts may be used in health foods for their immunomodulatory potential.
封面內頁
簽名頁
中文摘要 iii
英文摘要 iv
誌謝 v
目錄 vi
圖目錄 x
表目錄 xii

1.緒言 1
2.研究目的 4
3.文獻回顧 5
3.1巴蔘 5
3.2抗氧化 5
3.2.1自由基 5
3.2.2老化機制 7
3.2.3抗氧化物質 8
3.2.4抗氧化防禦系統 8
3.3免疫反應 11
3.3.1先天或非專一性免疫防禦 12
3.3.2後天或專一性免疫防禦 12
3.3.3免疫系統的細胞 13
3.3.4細胞激素 14
3.4癌症及免疫系統 26
3.4.1人類白血病細胞株U937 28
3.5一氧化氮 30
4.材料與方法 32
4.1實驗材料 32
4.1.1原料 32
4.1.2巴蔘萃取物之製備 32
4.1.3藥品 36
4.1.4儀器 37
4.2 實驗方法 38
4.2.1 抗氧化性分析 38
4.3 細胞試驗 40
4.3.1 U937白血病細胞之冷凍保存方法 40
4.3.2 U937白血病細胞之解凍方法 41
4.3.3 U937白血病細胞之培養方法 41
4.3.4活細胞計數法 41
4.3.5欲測試樣品之前處理 42
4.3.6人類單核球細胞條件培養液製備 42
4.3.7單核球細胞條件培養液間接抑制U937白血病細胞生長之試驗 43
4.4巴蔘萃取物經由間接模式對U937細胞抑制作用實驗 44
4.4.1人類單核細胞條件培養液製備 44
4.4.2樣品-單核細胞條件培養液間接抑制 U937 細胞生長之試驗 45
4.4.3樣品-單核細胞條件培養液細胞激素之含量測定 45
4.4.4測定經由樣品刺激後人類單核細胞之生長比例 47
4.4.5 NO檢測 49
4.4.6統計分析 49
5.結果與討論 50
5.1抗氧化性分析 50
5.1.1不同萃取溶劑之巴蔘葉及巴蔘莖萃取物之亞鐵離子螯合能力 50
5.1.2不同溶劑萃取之巴蔘葉及莖萃取物之DPPH自由基清除能力 53
5.1.3不同溶劑萃取之巴蔘葉及巴蔘莖萃取物之還原力 56
5.1.4不同溶劑萃取之巴蔘葉及巴蔘莖萃取物之Trolox當量抗氧化力 56
5.2利用熱水萃取之巴蔘葉及巴蔘莖萃取物間接抑制U937細胞生長之實驗 64
5.3利用熱水萃取之巴蔘葉及巴蔘莖萃取物促進人類單核細胞之生長效果 66
5.4條件培養液中細胞激素含量之測定 66
5.4.1條件培養液中細胞激素Interleukin-1β (IL-1β)之含量 68
5.4.2條件培養液中細胞激素Tumor necrosis factor-α (TNF-α)之含量 68
5.4.3條件培養液中細胞激素Interferon-γ(IFN-γ)之含量 71
5.5利用熱水萃取之巴蔘葉及巴蔘莖萃取物促進人類單核細胞條件培養液中一氧化氮(NO)之含量 73
5.6葉與莖及其萃取物之抗氧化分析 73
5.7葉與莖及其熱水萃取物之間接抑制U937細胞生長及 免疫調節之效果 76
6.結論 79
參考文獻 80


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