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研究生:郭宇薇
研究生(外文):Yu-Wei Guo
論文名稱:黃豆皂素粗萃取物對於1,2-dimethylhydrazine誘導F344大鼠結腸癌前期病變異常腺窩灶的影響及機制之探討
論文名稱(外文):The effect and mechanism of extracted crude soybean saponins on 1,2-dimethylhydrazine-induced ACF in F344 rats colon cancer
指導教授:林士祥
指導教授(外文):Shyh-Hsiang Lin
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:保健營養學系
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:2007
畢業學年度:95
語文別:中文
論文頁數:89
中文關鍵詞:異常腺窩灶黃豆皂素環氧化酶前列腺素β-葡萄糖醛酸酵素
外文關鍵詞:aberrant crypt foci (ACF)soybean saponinsprostaglandins (PGE2)cyclooxygenase (COX-2)β-glucuronidase
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本研究目的為探討黃豆皂素粗萃取物對結腸癌前期病變異常腺窩灶 (aberrant crypt foci, ACF) 的影響,並探討黃豆皂素粗萃取物降低結腸癌發生的可能機制。以雄性Fisher 344鼠做為動物模式,實驗分成五組在飲食當中添加不同劑量黃豆皂素粗萃取物,給予實驗飲食後第5週,以腹腔注射方式給予致癌物1,2-dimethylhydrazine (DMH) 誘發老鼠結腸癌的發生。在第15週予以犧牲,取出結腸,觀察結腸黏膜上ACFs數量、環氧化酶-2 (cyclooxygenase-2, COX-2) 蛋白質表現量、前列腺素E2 (prostaglandins E2, PGE2) 含量及β-葡萄糖醛酸酵素 (β-glucuronidase) 活性。結果顯示飲食中黃豆皂素粗萃取物可降低結腸黏膜上ACFs數量及β-glucuronidase活性,但不會影響COX-2蛋白質表現量及PGE2含量。飲食中的黃豆皂素粗萃取物對於結腸癌前期病變ACFs確實有一抑制功效,其機制可能與降低β-glucuronidase活性相關,而與COX-2及PGE2發炎反應的路徑無關。
This study investigated the effect of extracted crude soybean saponins on preneoplastic lesions, aberrant crypt foci (ACF), and the related inhibitory mechanism of extracted crude soybean saponins. Rats assigned into five groups according to different doses of extracted crude soybean saponins and received 1,2-dimethylhydrazine (DMH) injection in week 5. In week 15, all rats were sacrificed. The number of ACFs, the cyclooxygenase-2 (COX-2) protein expression, the level of prostaglandins E2 (PGE2) and the activity of β-glucuronidase were examed. Results revealed that the consumption of extracted crude soybean saponins decreased the number of ACFs and the activity of β-glucuronidase in rats, while did not affect the COX-2 protein expression and PGE2 level. In conclusion, extracted crude soybean saponins decreased the number and size of ACFs. The mechanism may be correlated to the activity of β-glucuronidase in colonic mucosa but not the COX-2 protein expression and PGE2 level.
目錄
中文摘要 I
Abstract II
致謝 III
目錄 V
圖目次 VIII
表目次 X
第一章 研究動機與目的 1
第二章 文獻回顧 3
第一節 結腸癌 3
(一) 腺窩與結腸癌 3
(二) 結腸癌之影響因子 4
(三) 結腸癌與發炎反應 7
(四) 結腸癌與腸道微生物酵素β-glucuronidase 11
(五) 結腸癌之動物模式 12
第二節 黃豆、黃豆皂素與異黃酮素 13
(一) 黃豆 13
(二) 黃豆皂素 13
(三) 異黃酮素 14
第三節 黃豆皂素之抗腫瘤作用 16
第三章 實驗設計與材料方法 18
第一節 實驗設計 18
(一) 實驗對象和飼養方式 18
(二) 實驗分組 18
(三) 實驗流程 19
(四) 劑量擬定方式 20
第二節 實驗材料 21
(一) 飼料成分 21
(二) 黃豆皂素粗萃取物製備 21
(三) 1, 2-dimethylhydrazine配製 21
第三節 分析項目及方法 23
(一) 動物犧牲及樣品收集 23
(二) 黃豆皂素粗萃取物成分分析 24
(三) 血漿異黃酮素分析 25
(四) 血脂分析 26
(五) Aberrant crypt foci染色觀察 26
(六) 老鼠結腸黏膜組織中COX-2蛋白質表現量 27
(七) 老鼠結腸黏膜組織中PGE2含量 30
(八) β-glucuronidase酵素活性測定 30
(九) 肝臟組織切片 31
第四節 試劑配方 32
第五節 統計方法 34
第四章 實驗結果 35
第一節 黃豆皂素粗萃取物分析 35
第二節 動物攝食和生長狀況 38
第三節 結腸癌相關指標 46
第五章 討論 55
第一節 黃豆皂素粗萃取物抗癌的成分 55
第二節 結腸癌的動物誘發模式 58
第三節 黃豆皂素抗癌功效 60
第四節 黃豆皂素對發炎反應相關物質的影響 62
第五節 黃豆皂素對腸道微生物酵素活性的影響 64
第六章 結論 65
第七章 參考文獻 66
附錄一 實驗試劑耗材與儀器一覽表 79
附錄二 動物與人體每公斤體重計量折算係數表 82
附錄三 SAS程式碼 83

圖目次
第二章
圖2-1、結直腸癌的發展 3
圖2-2、結腸癌誘發致癌物AOM對基因突變和蛋白質表現影響 10
圖2-3、不同種類黃豆皂素之化學結構式 14
圖2-4、不同種類之異黃酮素化學結構式 15

第三章
圖3-1、實驗分組 18
圖3-2、實驗流程 19
圖3-3、ACFs分類 27

第四章
圖4-1、利用HPLC觀察在波長248 nm下異黃酮素含量的層析曲線圖譜 36
圖4-2、利用HPLC觀察在波長260 nm下皂素含量的層析曲線圖譜 37
圖4-3、實驗期間老鼠體重變化 40
圖4-4、以Hematoxylin and eosin染色觀察老鼠肝臟組織切片 45
圖4-5、以亞甲藍染色觀察老鼠結腸黏膜上ACFs (10×40) 48
圖4-6、老鼠結腸黏膜COX-2蛋白質表現量 52
圖4-7、結腸黏膜PGE2濃度 53
圖4-8、結腸黏膜β-glucuronidase酵素活性 54






















表目次
第三章
表3-1、各組實驗飲食組成 22
表3-2、皂素HPLC分析移動相給予梯度濃度條件 24
表3-3、異黃酮素HPLC分析移動相給予梯度濃度條件 25

第四章
表4-1、老鼠體重增加、飼料攝食量及攝食效率 41
表4-2、老鼠肝臟重量及相對肝重 42
表4-3、老鼠結腸重量及相對結腸重 43
表4-4、老鼠第0及15週血脂分析 44
表4-5、老鼠小型ACFs計數 49
表4-6、老鼠大型的ACFs計數 50
表4-7、老鼠各段結腸ACFs計數 51
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