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研究生:周晧瑋
研究生(外文):How-Wei Chou
論文名稱:黑大蒜促使胃癌細胞凋亡及其作用機制之探討
論文名稱(外文):The mechanism of black garlic-induced apoptosis in human gastric cancer cell lines
指導教授:曾慶瀛曾慶瀛引用關係吳思敬
指導教授(外文):Chin-Yin TsengShe-Ching Wu
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:食品科學系研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:73
中文關鍵詞:黑大蒜人類胃癌細胞生物轉換細胞凋亡
外文關鍵詞:Black garlichuman gastric cancer cell (AGS)Biotransformationapoptosis
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從文獻當中指出:食用大蒜能顯著的降低胃癌的發生率。本實驗係採用經生物轉換之黑大蒜(Black garlic),利用其經過生物轉換(Bioconversion)之後,具有較強的抗氧化物活性、以及較高的硫化物成份含量等特性,進一步探討其殺死人類胃癌細胞(AGS)的效果,並藉由細胞凋亡因子之分析,來探討黑大蒜(Black Garlic)對於促使細胞凋亡之機制。
實驗結果顯示,藉由MTT分析法的結果,將人類胃癌細胞培養於濃度1.0 mg/ml的黑大蒜水萃液(BGWE)中,可達到抑制AGS細胞生長5 0 %;透過細胞計數法,相同濃度可造成1.5×105個AGS細胞死亡。利用Western blot分析BGWE對於AGS細胞凋亡相關蛋白的表現,發現當BGWE濃度為1.0 mg/ml時,細胞內凋亡因子AIF、cytochrome c、caspase-3蛋白表現有增加的趨勢,此外Bcl-2家族中的促進細胞凋亡因子Bax、Bid以及Bid上游調控因子FasL等蛋白質在細胞內也都有增加的表現。綜合以上結果, BGWE對AGS具有誘導細胞凋亡的作用。
According to the documents of prior researches, it was discovered that eating garlic noticeably induces the possibility of gastric cancer. This experiment focuses on the unique characteristics of black garlic. Through the process of bioconversion, black garlic shows stronger anti-oxidation and higher percentage of organic sulfur compounds, which enhances the effectiveness in killing human gastric cancer cell (AGS). Moreover through the analysis the factor of apoptosis, this experiment further discusses the role Black Garlic plays in the mechanism of apoptosis.The result of MTT analysis shows that by incubated AGS in 1.0mg/ml of BGWE, 50% of the AGS was eliminated; calculating by cell count, the same concentration of the extraction is capable to eliminate 1.5 X 105 of AGS. Western Blot was used to further analysis the efficacy of BGWE against AGS. It was discovered that when the concentration of BGWE is 1.0 mg/ml, the factor of apoptosis, AIF, cytochrome C, caspase-3 shows obvious growth in protein. As for Bcl-2 related BAX, Bid, …also indicates clear growth in protein.
Summarizing the above result, BGWE proves its capability in killing AGS and causes apoptosis.
目錄…………………………………………………………………..…Ⅰ
圖次…………………………………………………………………..…Ⅳ
表次…………………………………………………………………..…Ⅴ
摘要……………………………………………………………………ⅥI
Abstract…………………………………………………………….…ⅥII
前言………………………………………………………............……..IX
研究目的………………………………………………………………...X
研究架構….……………………………………………………..…….. XI
第一章 文獻回顧………………………………………………………..1
一、大蒜(Garlic)………………………………………………………2
(一) 大蒜的化學成分.........................................................................3
(二) 大蒜之抗腫瘤功效.....................................................................4
二、黑大蒜(Black garlic) .......................................................................6
(一)簡介...............................................................................................6
(二)黑大蒜之成分及功能性分析.......................................................7
三、胃癌簡介(Gastric cancer) .............................................................8
四、細胞凋亡(Apoptosis) ..................................................................11
第二章 材料及方法................................................................................19
一、實驗材料.....................................................................................20
(一)細胞...........................................................................................20
(二)黑大蒜.......................................................................................20
(三)藥品試劑...................................................................................20
(四)常用儀器...................................................................................22
二、實驗方法....................................................................................24
(一)細胞解凍培養...........................................................................24
(二)黑大蒜萃取物............................................................................25
(三)細胞數之計算............................................................................25
(四)細胞存活率試驗........................................................................26
(五)細胞毒性分析............................................................................27
(六) Annexin V-PI 染色法...............................................................28
(七)細胞週期分析............................................................................29
(八)西方點墨法................................................................................30
(九)統計分析....................................................................................33
第三章 結果與討論................................................................................34
一、黑大蒜萃取物(BGWE)毒殺胃癌細胞(AGS)之分析............................................................................................34
二、黑大蒜萃取物(BGWE)促使胃癌細胞(AGS)細胞死亡路徑之探討...........................................................................................39
(一) Annexin V-PI染色法之分析....................................................39
(二)細胞生長週期之分析................................................................39
(三)LDH release assay......................................................................41
三、黑大蒜萃取物(BGWE)對於胃癌細胞(AGS)細胞凋亡相關蛋白之分析...................................................................................45
四、結論............................................................................................55
第四章 參考文獻....................................................................................56










圖 次

圖1、細胞凋亡與細胞壞死之比較…………………………….…..…15
圖2、細胞凋亡途徑及其參與分子………………………….....…..…18















表 次

表1、大蒜之一般成分及含硫化合物含量....………………….…..…..3
表2、大蒜含硫化合物及其抗癌特性……………………….....…..…….5
表3、大蒜與黑大蒜之硫化物成分含量…………………….....…..…….7
表4、民國97年ICD-10全國主要癌症死亡原因………….....…..…….9
表5、細胞凋亡與細胞壞死之差異…………………………...…..…….16
表6、以MTT測試不同濃度之黑大蒜萃取物對人類胃癌細胞的存活率之影響……………………………………..……...….....…..…37
表7、不同濃度之黑大蒜萃取物對人類胃癌細胞的毒殺效果…………………….....................................................…..…….38
表8、以Annexin V-PI染色法分析不同濃度之黑大蒜萃取物對人類胃癌細胞死亡路徑之影響…………………….....………………...42
表9、分析不同濃度之黑大蒜萃取物對人類胃癌細胞生長週期之影響……….....……………………………………………………...43
表10、不同濃度之黑大蒜萃取物刺激人類胃癌細胞釋出乳酸脫氫酶比例之分析………….....................................................…..…….44
表11、分析不同濃度之黑大蒜萃取物對人類胃癌細胞Bax之表現………………….....…..……………………………………….49
表12、分析不同濃度之黑大蒜萃取物對人類胃癌細胞Bid之表現………………….....…..……………………..………………...50
表13、分析不同濃度之黑大蒜萃取物對人類胃癌細胞AIF之表現………………….....…..……………………..………………...51
表14、分析不同濃度之黑大蒜萃取物對人類胃癌細胞Cytochrome c之表現……………….....…..……………………..………………...52
表15、分析不同濃度之黑大蒜萃取物對人類胃癌細胞Caspase 3之表現………………….....…..……………………..………………...53

表16、分析不同濃度之黑大蒜萃取物對人類胃癌細胞FasL之表現………………….....…..……………………..………………...54
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