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  • 學位論文

數株Saccharomyces cerevisiae 細胞壁粗區分物之抗發炎及抑制腫瘤細胞增生功效研究

Anti-inflammatory activity and Anti-proliferation of tumor cells by the crude cell wall fractions of several Saccharomyces cerevisiae strains

指導教授 : 游若篍

摘要


發炎反應為人體受傷或抵禦外來微生物時之重要免疫反應,當人體處在發炎狀態時,一些發炎相關因子將會大量生成,若過度表現對人體為一種傷害。根據行政院衛生署統計資料顯示,2012 年十大癌症死因中,大腸癌及口腔癌分別佔據第三名和第五名。酵母菌的應用廣 泛,日常生活中常用來發酵啤酒等大宗發酵產品。本研究目的係以本實驗室所保有的五株S.cerevisiae 為材料,將其進行高溫高壓蒸汽蒸煮後,分別進一步製備成酵母菌細胞壁粗區分物crude mannoprotein (CM) 和crude β-glucan (CB),於LPS 誘導RAW 264.7 巨噬細胞模式之下,是否具有降低發炎情形,及抑制腫瘤細胞HSC-3、HT-29 增生的能力。結果顯示,S.cerevisiae BCRC 21447 之CM 在濃度1000 μg/ml 時,降低LPS 誘導RAW 264.7 分泌NO 含 量為五株酵母菌最多,相對LPS 組下降14.79%,而CB 在200 μg/ml 時,五株酵母菌皆有顯著性降低NO 含量( p < 0.05 ),其中又以S.cerevisiae BCRC 1599 ( 21.27% ) 效果最佳。此外CM 在濃度1000 μg/ml 時,具有較佳的清除DPPH 和NO 之能力,於西方墨點法的結果顯示,S. cerevisiae BCRC 21812 和S. cerevisiae BCRC 21447 之CB 可明顯隨著濃度增加而降低iNOS 蛋白質表現量,此外,五株酵母菌之CM 在濃度為125μg/ml 時能降低TNF-α 含量達顯著性差異( p < 0.05 ),而CB 在200 μg/ml 濃度下均顯著降低TNF-α 和IL-6 之生成量,在濃度100 μg/ml 以上時,也均顯著降低IL-1β 之生成量。最後,五株酵母菌細胞壁粗區分物CM和CB 對於兩株腫瘤細胞HSC-3、HT-29 具有抑制增生功效,如1000 μg/ml 的S.cerevisiae BCRC 21447 之CM 處理HSC-3 cell ,其存活率為60.94%;200 μg/ml 的CB 為70.42%。而 1000 μg/ml 的S. cerevisiae BCRC 21447 之CM 處理HT-29 cell ,其存活率為73.52%;200μg/ml 的CB 為75.49%。綜合上述結果,酵母菌細胞壁粗區分物CM 和CB 具有抗發炎和抑制HSC-3 ( oral ) and HT-29 ( colon ) 這兩株腫瘤細胞增生之功效。

並列摘要


Inflammation is an important human immune response against foreign microorganisms. When the body in a state of inflammation will generate some inflammatory factors. Department of Health (2013) statistics showed that cancer has been the first reason causes of death. Colon cancer and oral cancer occupy third place and five place of death of cancer in 2012. Saccharomyces cerevisiae is not only an excellent source of protein but also a functional food as probiotics. Another application of S.cerevisiae is used to ferment beer, wine or bread. The objective of this study was to investigate effects of crude cell wall fractions of several S. cerevisiae on LPS-induced RAW 264.7 macrophages. In addition, the DPPH, NO scavenging of crude cell wall fractions. Furthermore, the anti-proliferation of tumor cells of the crude cell wall fractions of several S. cerevisiae. The result revealed that both of crude mannoprotein (CM) and crude β-glucan (CB) suppressed production of NO. The highest inhibition ability followed by 1000 μg/ml CM from S. cerevisiae BCRC 21447 (85.21%, compare to LPS group) and 200 μg/ml CB from S. cerevisiae BCRC 1599 (78.73%, compare to LPS group). CM showed the better DPPH and NO scavenging activity than CB. Western blot analyses revealed that CB of S. cerevisiae inhibited protein expression of iNOS in LPS-induced macrophage; especially, S. cerevisiae BCRC 21812 and 21447. Also, 200 μg/ml CB of S. cerevisiae BCRC 21447 reduced TNF-α from 24.71 ± 1.09 ng/ml to 3.06 ± 0.15 ng/ml. All of the crude cell wall fractions lowered the production of IL -1β, IL-6 in high concentration. Moreover, these crude cell wall fractions of several S. cerevisiae showed various degree of the inhibitory effect on anti-proliferation of HSC-3, HT-29 tumor cells. 1000 μg/ml CM of S. cerevisiae BCRC 21447 inhibited survival rate of HSC-3 cells by 60.94%, 200 μg/ml CB inhibited survival rate of HSC-3 cells by 70.42%. 1000 μg/ml CM of S. cerevisiae BCRC 21447 inhibited survival rate of HT-29 cells by 73.52%, 200 μg/ml CB inhibited survival rate of HSC-3 cells by 75.49%. Conclusively, both CM and CB had anti-inflammatory activity and anti-proliferative activity of oral and colon cancer cell line.

參考文獻


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被引用紀錄


孫力勤(2016)。評估啤酒花萃取物的抗氧化與抗發炎活性〔碩士論文,國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342%2fNTU201601122

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