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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 游若? | |
dc.contributor.author | Shu-Ching Chiang | en |
dc.contributor.author | 江淑靜 | zh_TW |
dc.date.accessioned | 2021-06-15T04:54:46Z | - |
dc.date.available | 2015-08-10 | |
dc.date.copyright | 2010-08-10 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-29 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46125 | - |
dc.description.abstract | 益生質大多為非消化性寡醣,可通過胃酸及消化道而不被完全分解,到達大腸被腸道微生物利用。許多文獻指出,益生菌在利用非消化性寡醣發酵後所產生之二次代謝物和有機酸具有降低大腸癌細胞活性的功效。本研究欲評估四種寡醣 (包括菊醣、果寡醣、異麥芽寡醣和木寡醣) 對於乳酸桿菌和雙叉桿菌 (Lactobacillus casei 01、Bifidobacterium lactis Bb-12 和 Lactobacillus plantarum) 之助生性,並利用去除菌體之發酵液進行細胞試驗,觀察寡醣發酵液對於人類大腸癌細胞HT-29是否有抑制生長之效果,以及利用 4-nitroquiniline-1-oxide (4NQO) 誘導人類腸道細胞株 Int-407產生細胞和基因毒性,評估寡醣發酵液降低細胞和基因毒性的效果。最後分析寡醣發酵液之有機酸的組成,探討其組成和抑癌效果及抗基因毒性的關聯性。
從生長曲線和 pH 值變化發現,三珠菌株在添加不同寡醣之培養基中均可生長,除了菊醣添加之培養基的 pH 值變化並不顯著外,其它三種寡醣添加之培養基的pH值會隨著時間增加而降低。利用去除菌體後之寡醣發酵液與大腸癌細胞株 HT-29 共同培養 24 小時,除了 L. plantarum 之異麥芽寡醣發酵液無法抑制 HT-29 細胞株生長外,其它發酵液均有抑制 HT-29 細胞生長的效果 (p < 0.05),又以木寡醣的發酵液抑制癌細胞增生效果最好。而在抑制 4NQO 誘導之細胞毒性的實驗中,4NQO 組的細胞致死率約在 54% ,在添加寡醣發酵液共同培養後,多數寡醣發酵液均可顯著性提升細胞株的存活率 (p < 0.05)。在彗星電泳的顯影發現,添加寡醣發酵液的組別其 DNA 拖尾程度顯著性小於僅添加 4NQO 之細胞的 DNA 拖尾,平均又以B. lactis Bb-12之果寡醣發酵液效果較好。分析有機酸之含量發現,平均而言木寡醣發酵液的總短鏈脂肪酸濃度較高,推論可能與其抑制癌細胞能力有關;果寡醣發酵液中之丁酸濃度平均較其它寡醣發酵液高 (p < 0.05),推論可能與其抗細胞毒性與抗基因毒性的能力有關。 | zh_TW |
dc.description.abstract | Prebiotics are usually non-digestible oligosaccharides to human. They are resistant to gastric acid and digestive enzymes so that they could reach into the large intestines and were fermented by the intestinal microbes. Recently, some research has focused on the functions of the metabolites and short chain fatty acids that they might suppress the proliferation of colon adenocarcinoma cells. The aim of this study was to evaluate the anticytotoxic and antigentoxic effects of fermented supernatant by Lactobacillus casei 01, Bifidobaterium lactis Bb-12 and Lactobacillus plantarum in inulin, fructooligosacchardie (FOS), isomaltooligosaccharide (IMO) and xylooligosaccharide (XOS) contained medium.
The growth curves showed that three strains of probiotics could grow in the oligosaccharides-contained medium and the pH values were decreasing during the fermentation. The results showed that the fermented supernatant could suppress the proliferation of HT-29 (p < 0.05) except L. plantarum-IMO group. In addition, the XOS-fermented supernatant showed the best suppression. The viability of 4NQO group was 54% and almost of cell viability in the test groups increased by treating with the fermented supernatant (p < 0.05), and the FOS-fermented supernatant showed the best ability in the promoting the cell viability after 4NQO induced cell damages. Furthermore, we observed that the DNA tails reduced in the test groups significantly by comet assay (p < 0.05). From the analysis of organic acids, the XOS-fermented supernatant contained higher concentration of total SCFAs that might contribute to the decreasing viabilities on HT-29. FOS-fermented supernatant contained the highest butyrate concentration which might contribute to the anticytotoxicity and antigenotoxicity. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:54:46Z (GMT). No. of bitstreams: 1 ntu-99-R97641032-1.pdf: 1417224 bytes, checksum: 43a89ca497c3feca2404aa09268405d8 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 目錄
謝誌 i 摘要 iii 英文摘要 iv 目錄 v 圖目錄 ix 表目錄 xi 壹、前言 1 貳、文獻整理 2 一、人類腸道微生物的功能與健康 2 (一) 保護和結構性功能 2 (二) 營養性功能 3 (三) 代謝性功能 3 二、益生質(Prebiotics) 5 (一) 益生質的訂定標準 5 (二) 益生質的消化代謝 8 (三) 食品工業之應用 10 (四) 益生質的種類與製造: 13 1.菊醣類寡醣 13 2.異麥芽寡醣 13 3.木寡醣 14 三、益生質對健康的功效 16 (一) 影響腸胃道微生物菌相 16 (二) 改善陰道感染 16 (三) 對於第二型糖尿病和肥胖者的益處 16 (四) 改善腹瀉 17 (五) 改善便秘 17 (六) 降低新生兒罹患腸壞死症的機率 18 (七) 影響礦物質吸收 18 (八) 降低心血管疾病 18 四、大腸癌 19 (一) 癌症的發生 19 (二) 致癌物質 22 五、益生質、益生菌 (Probiotics)和大腸癌 (Colorectal cancer)的相關性 24 (一) 抑制突變物以及抗基因毒性的活性 24 (二) 去毒性作用 (detoxification) 25 (三) 調節酵素系統使致癌物失活 25 (四) 健全腸道障壁系統 25 (五) 發酵產物和乳酸菌之二次代謝物 26 參、材料與方法 28 一、實驗架構 28 二、實驗材料 30 (一) 試驗寡醣 30 (二) 試驗菌株 30 (三) 試驗細胞株 30 (四) 藥品與培養基 30 (五) 器材 31 三、實驗方法 33 (一) 菌種的保存與活化 33 (二) 乳酸菌於寡醣添加之培養基的助生試驗與pH變化 33 (三) 發酵液的製備 34 (四) 細胞株的活化、繼代培養與保存 34 (五) MTT assay 36 (六) 益生菌之寡醣發酵液抑制人類大腸癌細胞株 HT-29 試驗 36 (七) 益生菌之寡醣發酵液對人類腸道細胞 Int-407 存活率影響試驗 37 (八) 乳酸菌之寡醣發酵液抑制 4NQO 誘導 Int-407 之細胞毒性試驗 37 (九) 彗星電泳 (Comet assay) 38 (十) 乳酸菌之寡醣發酵液抑制 4NQO 對 Int-407 細胞毒性之機制探討 39 (十一) 發酵液之有機酸分析 41 四、統計分析 42 肆、結果與討論 43 一、四種益菌助生寡醣對於 L. casei 01 、 B. lactis Bb-12 和 L. plantarum 生長的影響其發酵期間的pH變化 43 (1) 四種益菌助生寡醣對於 L. casei 01 生長的影響及 pH 變化 43 (2) 四種益菌助生寡醣對於 B. lactis Bb-12 生長的影響及 pH 變化 44 (3) 四種益菌助生寡醣對於 L. plantarum 生長的影響及 pH 變化 45 二、益生菌的寡醣發酵液對於人類大腸癌細胞 HT-29 的抑制效果 46 三、益生菌之寡醣發酵液對人類腸道細胞株之毒性評估試驗 48 四、寡醣之發酵產物抑制 4NQO 誘導之細胞毒性試驗 48 五、寡醣之發酵產物抑制 4NQO 誘導之基因毒性試驗 49 六、寡醣之發酵產物抑制 4NQO 誘導細胞毒性之機制探討 50 (1) 寡醣發酵液對 4NQO 之阻斷效應 (Blocking effects) 50 (2) 寡醣發酵液對 4NQO 之去致突變效應 (Destoxic effects) 51 (3) 寡醣發酵液對 4NQO 之生物抗致突變效應 (Bioantitoxic effects) 51 七、三株乳酸菌於四種寡醣添加之培養基中進行發酵後之有機酸分析 52 伍、結論 56 陸、參考文獻 錯誤! 尚未定義書籤。 附錄 錯誤! 尚未定義書籤。 附圖一、Int-407細胞株DNA損傷之顯微照片 錯誤! 尚未定義書籤。 附表一、益生菌之寡醣發酵液與HT-29細胞增生之影響 錯誤! 尚未定義書籤。 附表二、益生菌之寡醣發酵液對Int-407細胞株之毒性試驗 錯誤! 尚未定義書籤。 附表三、益生菌之寡醣發酵液對4NQO誘導腸道細胞Int-407細胞毒性之影響 錯誤! 尚未定義書籤。 附表四、益生菌之寡醣發酵液抗4NQO誘導之細胞毒性的機制探討 錯誤! 尚未定義書籤。 遵照台灣農業化學與食品科學期刊之稿約撰寫 58 圖目錄 圖 一、腸內微生物的作用 4 圖 二、腸道微生物利用非消化性寡醣之代謝圖 9 圖 三、非消化性寡醣的製造流程 15 圖 四、癌細胞產生的階段及抑癌物質在不同階段的作用 21 圖 五 (a,b)、(a) 4NQO 與鳥嘌呤鍵結的結構 (b) 4NQO 及其代謝衍生物 23 圖 六、短鏈脂肪酸對腸道細胞癌化過程中的影響 27 圖 七、實驗架構 29 圖 八、 L. casei 01 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的生長情形 58 圖 九、 L. casei 01 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的 pH 變化 59 圖 十、 B. lactis Bb-12 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的生長情形 60 圖 十一、 B. lactis Bb-12 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的 pH 變化 61 圖 十二、 L. plantarum 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的生長情形 62 圖 十三、 L. plantarum 在含有菊醣、果寡醣、異麥芽寡醣和木寡醣之培養基中的 pH 變化 63 圖 十四、 L. casei 01 之發酵產物對於 HT-29 細胞株增生之影響 64 圖 十五、 B. lactis Bb-12 之發酵產物對於 HT-29 細胞株增生之影響 65 圖 十六、 L. plantarum 之發酵產物對於 HT-29 細胞株增生之影響 66 圖 十七、寡醣發酵液對 Int-407 細胞株之毒性試驗 67 圖 十八、 L. casei 01 之發酵產物對 4NQO 誘導腸道細胞 Int-407 細胞毒性之影響 68 圖 十九、 B. lactis Bb-12 之發酵產物對 4NQO 誘導腸道細胞 Int-407 細胞毒性之影響 69 圖 二十、L. plantarum之發酵產物對4NQO誘導腸道細胞Int-407細胞毒性之影響 70 圖 二十一、寡醣發酵液對 4NQO 誘導 Int-407 之細胞毒性的阻斷效應 71 圖 二十二、寡醣發酵液對 4NQO 誘導 Int-407 細胞毒性之去致突變效應 72 圖 二十三、寡醣發酵液對 4NQO 誘導 Int-407 細胞毒性之生物抗致突變效應 73 表目錄 表一、數種寡醣的益生性評估 7 表二、常見的功能性寡醣 12 表三、寡醣發酵液對4NQO誘導Int-407細胞株DNA損傷之影響 74 表四、接種L. casei 01與四種寡醣進行發酵24小時後之有機酸成分分析 75 表五、接種B. lactis Bb-12與四種寡醣進行發酵24小時後之有機酸成分分析 76 表六、接種L. plantarum與四種寡醣進行發酵24小時後之有機酸成分分析 77 | |
dc.language.iso | zh-TW | |
dc.title | 益生菌寡醣發酵液抑制大腸癌細胞增生及其對人類結腸細胞株Int-407抗細胞毒性研究 | zh_TW |
dc.title | Effects of Probiotcis-Fermented Supernatant from Substrates Containing Oligosaccharides on the Inhibition of Colon Cancer Cell Proliferation and Antitoxicity of Human Colon Cell Line Int-407 | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 周正俊,丘志威,潘崇良,蔡國珍 | |
dc.subject.keyword | 寡醣,發酵,短鏈脂肪酸,大腸癌,抗細胞毒性, | zh_TW |
dc.subject.keyword | oligosaccharides,fermentation,short chain fatty aicds,colon cancer,anticytotoxicity, | en |
dc.relation.page | 93 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-07-30 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 食品科技研究所 | zh_TW |
顯示於系所單位: | 食品科技研究所 |
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