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研究生:周邦彥
研究生(外文):BANG-YAN Zhou
論文名稱:南非葉萃取物對於人類癌細胞的增殖抑制效果
論文名稱(外文):Inhibition of human cancer proliferation by the extracts of Vernonia amygdalina
指導教授:蘇文達蘇文達引用關係
指導教授(外文):Wen-Ta Su
口試委員:吳漢忠柯智升陳文章
口試日期:2016-07-22
學位類別:碩士
校院名稱:國立臺北科技大學
系所名稱:化學工程與生物科技系生化與生醫工程碩士班
學門:工程學門
學類:化學工程學類
論文種類:學術論文
畢業學年度:104
中文關鍵詞:南非葉、抗氧化活性、超臨界流體萃取、HCT116、SKHep、抗癌
外文關鍵詞:Vernonia amygdalinaantioxidant activitysupercritical fluid extractionHCT116SKHep-1anticancer
相關次數:
  • 被引用被引用:1
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  • 下載下載:24
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本研究的目的是使用超臨界流體二氧化碳(SC-CO 2)的效果提取用於從南非葉得到的癌抑制化合物。超臨界流体萃取沒有溶劑殘留的問題,SC-CO 2萃取物比傳統的有機回流提取物有更快和更環保的方法。從萃取物SC-CO 2萃取的最佳條件為40兆帕,50℃,60分鐘的操作,並在最大收率為1.908%。從南非葉提取物的清除自由基的活性,還原力、ABTS和DPPH測定具有優異的抗氧化能力。南非葉提取物能有效抑制肝癌細胞(SKHEP-1)和結腸癌細胞(HCT116)細胞在體外實驗的生存率。SKHEP-1的IC50為332.6167±4.07 μg/ml,HCT116的IC50為559.5±42.5μg/ml。因此,南非葉通過超臨界CO2萃取方式抑制某些人類癌症有潛在能力。
The objective of this study was to investigate the effect of supercritical fluid CO2 (SC-CO2) extracts for the cancer suppression compound obtained from Vernonia amygdalina. SC-CO2 extraction is quicker and more environmental friendly method than traditional organic reflux extraction and there are absences of solvent residues in the extracts. The optimal condition of SC-CO2 extraction from Vernonia amygdalina was 40 MPa, 50°C, and 60 min operation, and the maximal yield was 1.908 %. The radical-scavenging activity of extract from Vernonia amygdalina was measured by reducing power, ABTS, and DPPH assays and these extracts possess very excellent antioxidant capacity. The extracts of Vernonia amygdalina can effectively inhibit the viability of hepatic cancer cell (SKHEP-1) and colon cancer cell (HCT116) cells in vitro. The IC50 value was 332.6167±4.07 μg/mL for SKHEP-1, and 559.5±42.5 μg/mL for HCT116. Therefore, the extracts of Vernonia amygdalina by supercritical fluid CO2 extraction have a potential curing ability for some human cancers.
摘 要 I
ABSTRACT II
誌謝 IV
目錄 V
表目錄 VIII
圖目錄 IX
第一章 緒論 1
1.3實驗架構 2
第二章 文獻回顧 3
2.1癌症 3
2.1.1 癌症 3
2.1.2癌細胞特徵 3
2.1.3 癌症凋亡 8
2.1.4 肝癌 10
2.1.5腸癌 12
2.2南非葉 13
2.2.1 起源以及命名 13
2.2.2傳統中草藥功用 14
2.3 萃取 15
2.3.1超臨界流體 15
2.3.2 乙醇萃取 17
2.4抗氧化能力 17
2.4.1 ABTS 18
2.4.2 DPPH 18
2.4.3 還原力 19
第三章 材料與方法 20
3.1 實驗材料、藥品與儀器 20
3.1.1材料與藥品 20
3.1.2 儀器設備型號 21
3.2實驗方法 22
3.2.1 南非業的前置處理 22
3.2.2萃取方式 22
3.2.2.1 迴流萃取萃取方式 22
3.2.2.2 超臨界萃取 22
3.2.2.3 產率 23
3.2.3抗氧化能力測試 23
3.2.3.1 DPPH 23
3.2.3.2 ATBS 23
3.2.3.3 還原力 24
3.2.4 細胞培養 24
3.2.5 細胞存活率分析(MTT) 25
3.2.6 藥效 25
3.2.7 細胞凋亡 26
3.2.8 細胞週期 26
3.2.9 免疫螢光染色法 27
3.2.10 GC-MS成分鑑定 28
第四章 實驗結果 29
4.1南非葉產率 29
4.1.1 萃取方式 29
4.1.2高壓極性超臨界流體參數調整 30
4.1.3低壓非極性超臨界流體參數調整 31
4.2抗氧化能力測試 32
4.2.1 DPPH 32
4.2.2 ABTS 33
4.2.3 REDUCING POWER 34
4.3 實驗小結論(一) 34
4.4 南非葉萃取方式對於存活率的影響 35
4.5 實驗小結論(二) 36
4.6 南非葉萃取物濃度對於細胞的影響 36
4.7 南非葉萃取物的藥效測試 39
4.8 實驗小結論(三) 40
4.9 南非葉萃取物對於癌細胞的細胞凋亡影響 40
4.10 南非葉萃取物對於癌細胞的細胞週期影響 42
4.11 南非葉萃取物對於癌細胞的BAX免疫蛋白染色 44
4.12南非葉萃取物對於癌細胞的BCL-2免疫蛋白染色 45
4.13 萃取物對於癌細胞的CASPASE3免疫蛋白染色 47
4.14 萃取物對於癌細胞的CASPACE9免疫蛋白染色 48
4.15 超臨界流體萃取高壓葉子萃取的GC-MS 50
第五章 結論 52
參考文獻 54
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