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| 題名: | 優酪乳對DMH誘發F344鼠大腸癌前期病變之預防效果
Preventive effects of yogurt on DMH-induced preneoplastic lesions of colon in F344 rats |
| 作者: | 林明萱
Lin Ming-Shiuan |
| 貢獻者: | 保健營養學研究所
施純光 |
| 關鍵詞: | 碩士
指導教授-施純光
委員-江文章
委員-商惠芳
大腸癌;優酪乳;盲腸內容物;異常腺窩病灶;黏液素;第二型環氧合酶;免疫球蛋白A
Colorectal cancer;Yogurt;Cecal contents;Aberrant crypt foci;Mucin;Cyclooxygenase-2;Immunoglobulin A |
| 日期: | 2010 |
| 上傳時間: | 2010-10-20 12:11:14 (UTC+8) |
| 摘要: | 惡性腫瘤位居台灣十大死因之首，其中結腸直腸癌(大腸癌)在癌症死因中排名第三位，飲食與大腸癌發生最具關聯性。優酪乳是以牛奶為基礎，用益生菌作為發酵菌而製成的產品，先前研究發現牛奶及優酪乳對於大腸癌可能有預防的效果。本研究目的為評估原料乳或以不同菌株發酵之優酪乳是否具有預防大腸癌發展的功效。利用化學致癌劑誘發的大腸癌動物模式，以蒸餾水(負控制組)、化學預防劑Piroxicam(正控制組)及優酪乳製造過程中的原料乳、基礎優酪乳(Streptococcus thermophilus及Lactobacillus bulgaricus)、含A菌優酪乳(S. thermophilus、L. bulgaricus和L. acidophilus)、含B菌優酪乳(S. thermophilus、L. bulgaricus和Bifidobacterium lactis)與含A、B菌優酪乳(S. thermophilus、L. bulgaricus、L. acidophilus和B. lactis)對F344大鼠施行管餵，在實驗飲食介入一週後以腹腔注射給予1,2-Dimethylhydrazine (DMH)誘發大腸癌，十週後犧牲大鼠。取其盲腸內容物檢測pH值及β-glucuronidase活性；大腸檢測異常腺窩病灶(Aberrant crypt feci, ACF)、黏液素分泌型態，而大腸黏膜分析發炎性蛋白第二型環氧合酶、誘導型一氧化氮合成酶表現量；並測其血液中免疫球蛋白A濃度。盲腸內容物之pH值及β-glucuronidase活性在各組間都無顯著差異。正控制組、原料乳組、基礎發酵乳組、含A菌優酪乳組及同時含A、B菌優酪乳組其大腸之ACF及異常腺窩數顯著低於負控制組，且基礎優酪乳組、含A菌優酪乳組及同時含A、B菌優酪乳組遠端大腸分泌sialomucin之ACF數顯著低於負控制組，然而黏液素缺乏病灶在各組間都無顯著差異。含B菌優酪乳組及同時含A、B菌優酪乳組近端結腸COX-2蛋白質表現顯著高於負控制組。所有的優酪乳組別血清免疫球蛋白A濃度顯著低於負控制組。本研究顯示餵食基礎優酪乳、含A菌優酪乳及同時含A、B菌優酪乳連續十週對於大腸癌發展早期指標具顯著抑制效果，推測是藉由抑制大腸癌形成過程黏液素的變化，而達到預防大腸癌的效果。
Malignancy is the first leading cause of death in Taiwan. Colorectal cancer, the third cause of cancer death in Taiwan, is the form of cancer most associated with diet. Yogurt is produced based on milk and fermented by probiotics. Previous study showed that milk and yogurt could prevent colorectal cancer. The purpose of this study was to assess the preventive effect of milk and yogurts fermented by various probiotics on the development of colorectal cancer. F344 rats were fed distilled water (negative control), piroxicam (positive control), milk, bsal yogurt (Streptococcus thermophilus and Lactobacillus bulgaricus), yogurt A (S. thermophilus, L. bulgaricus and L. acidophilus), yogurt B (S. thermophilus, L. bulgaricus and Bifidobacterium lactis) or yogurt AB (S. thermophilus, L. bulgaricus, L. acidophilus and B. lactis). After one week of feeding, rats were injected intraperitoneally with 1,2-dimethylhydrazine to induce colon carcinogenesis. After 10 weeks of feeding, rats were sacrificed. The cecal contents were examined for pH and β-glucuronidase activity; the colons were examined for aberrant crypt foci (ACF), mucin production, and the colonic mucosa for pro-inflammatory cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) protein expression; the blood were were examined for imunoglobulin A (Ig A) levels. The results showed that there were no significant differences in cecal pH and β-glucuronidase activity among groups. Postive control, milk, basal yogurt, yogurt A and yogurt AB significantly reduced the numbers of ACF and aberrant crypts in the colon as compared to negative control. Basal yogurt, yogurt A and yogurt AB significantly reduced the number of sialomucin-producing ACF in the distal colon as compared to negative control. There were no significant differences in mucin-depleted foci (MDF) number among groups. Yogurt B and AB significantly increased the COX-2 protein expression of the proximal colon as compared to negative control. All yogurt groups significantly reduced the plasma Ig A levels as compared to negative control. This study showed that basal yogurt and yogurt A and yogurt AB significantly inhibited the early markers of colorectal cancer after 10-week feeding, suggesting that these yogurts may inhibit colon carcinogenesis via mucin modification. |
| 關聯: | 153頁 |
| 描述: | 目錄
中文摘要……………………………………………………………………………….I
英文摘要……………………………………………………………………………..III
目錄……………………………………………………………………………..…….V
圖目錄……………………………………………………………………………....VII
表目錄……………………………………………………………………………...VIII
第一章 前言…………………………………………………………………………..1
第二章 文獻回顧……………………………………………………………………..3
第一節大腸癌…………………………..……………………………………3
(一)大腸構造與功能………………………………..………………...3
(二)大腸癌簡介.……………………………………....………………5
(三)大腸癌的形成及分期…………………………………………….6
(四)大腸癌與ACF…………………………………………………….9
(五)大腸癌與黏液素………………………………………………...11
(六)大腸癌與發炎性蛋白質COX-2及iNOS………………………13
(七)大腸癌與糞便生理………………………………………...........17
(八)免疫球蛋白A與腸道免疫………………………………..........21
第二節 牛奶介紹…………………………………………………........22
（一） 牛奶簡介……………………………………….………….22
（二） 牛奶與大腸癌的研究…………………………….……….23
第三節 優酪乳介紹…………………………………...……………….25
（一） 菌種介紹…………………………………………………..25
（二） 優酪乳簡介………………………………………………..28
（三） 優酪乳之機能性研究……………………………………..30
（四） 優酪乳與大腸癌的研究…………………………………..36
第三章 研究目的……………………………………………………………………39
第四章 材料與方法……………………………………………………………........40
第一節 實驗材料……………………………..……………………..……..40
（一） 實驗動物…………………………………………………..40
（二） 飼料成分…………………………………………………..40
（三） 實驗藥品…………………………………………………..41
（四） 儀器設備…………………………………………………..44
（五） 樣品來源…………………………………………………..46
第二節 實驗分組與動物飼養……………………………………………..48
第三節 實驗流程…………………………………………………………..49
第四節 分析項目…………………………………………………………..50
（一） 盲腸內容物之pH值及β-glucuronidase活性分析……..50
（二） ACF分析………………………………………………….51
（三） 黏液素分析………………………………………………..52
（四） 黏膜發炎性蛋白分析……………………………………..53
（五） 血漿中Ig A分析………………………………………….55
第五節 統計分析…………………………………………………………..56
第五章 結果 ………………………………………………………………………..57
第一節 動物攝食、生長狀況及肝臟與腸道生理之影響………………..57
第二節 ACF分析………………………………………………………….58
第三節 ACF黏液素分泌型態及MDF……………………………………61
第四節 發炎性蛋白質表現…………………………..……………………62
第五節 血漿Ig A濃度…………………………………………………….63
第六節 大腸癌分析指標之相關性………………………………………..64
第六章 討論……………………………………………………………...………….65
第一節 優酪乳對動物攝食、生長狀況及肝臟與腸道生理之影響...……65
（一）優酪乳對動物攝食及生長狀況之影響...….………….…….65
（二）優酪乳對動物肝臟及腸道生理之影響...…….…………….65
第二節 優酪乳對ACF之影響……………………………………………68
第三節 優酪乳對ACF黏液素分泌型態及MDF之影響………………...74
第四節 優酪乳對發炎性蛋白質表現之影響……………………………..78
第五節 優酪乳對血漿Ig A濃度之影響……………………….…………80
第七章 結論………………………………………….……………………………...82
第八章 參考文獻……………………………………………………………………83
圖目錄
圖1、人體大腸及腸壁構造圖….……..…………….……………………………...116
圖2、正常大腸腺窩形態.......….………………………………………………….117
圖3、DMH的代謝過程.…………..…………….……………………………..........118
圖4、ACF外觀……………………………………………………………………119
圖5、胃腸道對於細菌刺激所產生非免疫學及免疫防禦…………….………...120
圖6、優酪乳製造過程……..…………….…………………………….................121
圖7、實驗流程圖……………..…………….………………………………….....122
圖8、實驗架構圖………………..…………….…………………………………..123
圖9、餵食優酪乳對雄性F344鼠盲腸內容物β-glucuronidase比活性之影響….124
圖10、以Methylene blue染色觀察DMH 誘發大鼠大腸組織顯示正常腺窩及ACF…………………………………………………………………………………125
圖11、結腸ACF黏液素染色圖……………………………….…………………...126
圖12、餵食優酪乳對雄性F344鼠近端結腸COX-2蛋白質表現量之影響……127
圖13、餵食優酪乳對雄性F344鼠大腸中間段大腸COX-2蛋白質表現量之影響……………………………………………………………………………………128
圖14、餵食優酪乳對雄性F344鼠大腸遠端大腸COX-2蛋白質表現量之影響..129
圖15、 ACF的生長透過分裂機制…….………………………………………….130
表目錄
表1、TNM分類………………..…………….………………………………..…..131
表2、腸癌之Dukes分類系統……………..…………….……………………..…132
表3、優酪乳之組成……………..…………….……………………………….…133
表4、優酪乳之營養標示………………..…………….………………………..…134
表5、優酪乳之乳酸桿菌數、雙岐桿菌數及總乳酸菌數……..…………………135
表6、實驗組別………………..…………….…………………………………..…136
表7、生長期老鼠AIN-93G標準飼料組成表.…………………………………..…137
表8、不同蛋白質之抗體及稀釋倍率………………………………………....…138
表9、 Ig A標準曲線……..…………….………………………………………..…139
表10、餵食優酪乳製品對雄性F344鼠體重、攝食量及餵食效率之影響….…...140
表11、餵食優酪乳製品對雄性F344鼠肝臟重量之影響…….……………….…141
表12、餵食優酪乳製品對雄性F344鼠盲腸內容物之影響….………………… 142
表13、餵食優酪乳製品對雄性F344鼠大腸長度及大腸重量之影響.………… 143
表14、餵食優酪乳製品對雄性F344 鼠大腸受DMH 所誘發ACF 之影響…….144
表15、餵食優酪乳製品對雄性F344 鼠大腸受DMH 所誘發各種大小ACF 之影響………….………………………………..…………….…………………………145
表16、餵食優酪乳製品對雄性F344 鼠大腸受DMH 所誘發ACF 分布之影響……………..…………….…………………………………..…………….…..…146
表17、餵食優酪乳製品對雄性F344 鼠大腸受DMH 所誘發各段大腸各種大小ACF 分布之影響…………..…………..…………….……………………...…..…147
表18、餵食優酪乳製品對雄性F344鼠遠端大腸受DMH所誘發各種黏液素類型ACF數之影響……………….…………………………………..…………….……148
表19、餵食優酪乳製品對雄性F344鼠遠端大腸受DMH所誘發各種黏液素類型ACF多發性之影響……………..…………..…………….………………….…..…149
表20、餵食優酪乳製品對雄性F344鼠大腸受DMH所誘發MDF之影響……..150
表21、餵食優酪乳製品對雄性F344 鼠大腸受DMH 所誘發MDF 分布之影響……………………………………………………………………………………151
表22、餵食優酪乳對雄性F344鼠血漿Ig A濃度之影響……………………....…152
表23、大腸癌各項指標之相關性………………….……………………………..153
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