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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 周正俊 | |
| dc.contributor.author | Chien-Li Liao | en |
| dc.contributor.author | 廖建俐 | zh_TW |
| dc.date.accessioned | 2021-06-13T03:46:54Z | - |
| dc.date.available | 2006-07-31 | |
| dc.date.copyright | 2006-07-31 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-26 | |
| dc.identifier.citation | 陸、參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32396 | - |
| dc.description.abstract | 本實驗以豆奶為基質，利用乳酸菌 (Streptococcus salivarius subsp. thermophilus CCRC 14085) 單獨或與雙叉桿菌 (Bificobacterium infantis CCRC 14633) 共同發酵，以 lipopolysaccharides (LPS) 誘導巨噬細胞 RAW 264.7之模式，探討豆奶及發酵豆奶甲醇萃取物對 nitric oxide (NO) 、 tumor necrosis factor (TNF)-α 、 interleukin (IL)-6 、 IL-1β 等發炎因子之影響，並進一步以Western blot分析巨噬細胞 inducible nitric oxide synthase (iNOS) 與 cyclooxygenase-2 (COX-2) 之蛋白質表現量。
結果顯示，豆奶及兩種發酵豆奶甲醇萃取物皆可有效抑制由 LPS 誘導所產生之 NO，且抑制能力隨著濃度增加而提升，其中又以豆奶甲醇萃取物之抑制能力最佳，其次為 S. thermophilus 發酵豆奶甲醇萃取物。iNOS酵素活性分析結果，推測此三種樣品均不改變 iNOS 酵素活性，而除豆奶甲醇萃取物之外， S. thermophilus 發酵豆奶與 S. thermophilus 及 B. infantis 共同發酵豆奶甲醇萃取物可明顯抑制經由 LPS 誘發產生之 iNOS 蛋白質表現量。另外，豆奶及兩種發酵豆奶甲醇萃取物不具抑制 COX-2 蛋白質表現之效用。於細胞激素試驗中發現，豆奶、 S. thermophilus 發酵豆奶與 S. thermophilus 及 B. infantis共同發酵豆奶甲醇萃取物均具有抑制 LPS 誘導 RAW 264.7巨噬細胞生成 TNF-α、 IL-6 及 IL-1β 之能力。 | zh_TW |
| dc.description.abstract | In the present study, the methanol extracts of soymilk and soymilk fermented with lactic acid bacteria (Streptococcus salivarius subsp. thermophilus CCRC 14085) alone or with bifidobacteria (Bificobacterium infantis CCRC 14633) simultaneously were evaluated for their effects on lipopolysaccharide (LPS)-induced nitric oxide, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β production by RAW 264.7 macrophages. In addition, the proteins expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by macrophages as influenced by these methanol extracts were also investigated.
Results revealed that the methanol extracts of soymilk and the prepared fermented soymilk dose-dependently suppress NO production of LPS-induced macrophages, with soymilk showed the highest inhibition ability followed by S. thermophilus-fermented soymilk. Extracts of soymilk and fermented soymilk were found not affected iNOS enzyme activity. Western blot analyses revealed that the methanol extracts of the fermented soymilk inhibited protein expression of iNOS in LPS-induced macrophages while it was not observed with soymilk. Moreover, these extracts did not show obvious inhibition on COX-2 protein expression in LPS-induced macrophages. On the other hand, the methanol extracts of soymilk and fermented soymilk all showed the inhibitory effect on LPS-induced TNF-α, IL-6 and IL-1β production by RAW 264.7 macrophages. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T03:46:54Z (GMT). No. of bitstreams: 1 ntu-95-R93641022-1.pdf: 2393073 bytes, checksum: bb6f1c81cef563377c9a668ebfcb4b23 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 目錄
中文摘要-------------------------------------------------------------------------- I Abstract-------------------------------------------------------------------------- II 目錄 ------------------------------------------------------------------------------- III 圖次 ------------------------------------------------------------------------------- VI 表次 ------------------------------------------------------------------------------- VII 壹、前言 ------------------------------------------------------------------------- 1 貳、文獻整理 ------------------------------------------------------------------- 3 （一）黃豆及其加工製品---------------------------------------------------- 3 一、黃豆------------------------------------------------------------------ 3 1. 黃豆簡介------------------------------------------------------------ 3 2. 黃豆生理活性成分------------------------------------------------ 3 二、異黃酮素------------------------------------------------------------ 4 1. 異黃酮素簡介------------------------------------------------------ 4 2. 異黃酮素之生理機能--------------------------------------------- 4 (1) 抗氧化活性----------------------------------------------------- 4 (2) 抗心血管疾病-------------------------------------------------- 6 (3) 預防骨質疏鬆症----------------------------------------------- 6 (4) 類雌激素及抗雌激素特性----------------------------------- 6 (5) 降低癌症罹患率----------------------------------------------- 7 三、豆奶與發酵豆奶---------------------------------------------------- 7 1. 豆奶------------------------------------------------------------------ 7 2. 發酵豆奶------------------------------------------------------------ 8 （二）益生菌------------------------------------------------------------------- 11 一、益生菌簡介--------------------------------------------------------- 11 1. 益生菌定義--------------------------------------------------------- 11 2. 益生菌應具備之條件--------------------------------------------- 12 3. 益生菌促進人體健康效益之方式------------------------------ 12 二、乳酸菌簡介--------------------------------------------------------- 14 三、雙叉桿菌簡介------------------------------------------------------ 14 （三）發炎反應與相關因子------------------------------------------------- 15 一、發炎反應與巨噬細胞---------------------------------------------- 15 二、LPS 與巨噬細胞之活化------------------------------------------ 15 三、NO 及 iNOS ------------------------------------------------------- 17 四、PGE2 及 COX-2---------------------------------------------------- 21 五、Pro-inflammatory cytokines -------------------------------------- 23 六、異黃酮素與發炎反應---------------------------------------------- 24 七、益生菌與發炎反應------------------------------------------------- 24 參、實驗架構------------------------------------------------------------------ 26 肆、材料與方法---------------------------------------------------------------- 27 （一）實驗材料---------------------------------------------------------------- 27 一、原料-------------------------------------------------------------------- 27 二、試驗菌種 -- 27 三、細胞株---------------------------------------------------------------- 27 四、培養基---------------------------------------------------------------- 27 五、化學藥品及試劑---------------------------------------------------- 27 （二）儀器設備---------------------------------------------------------------- 29 （三）實驗方法---------------------------------------------------------------- 30 一、樣品製備-------------------------------------------------------------- 30 1. 接種源之保存活化與製備--------------------------------------- 30 2. 豆奶基質------------------------------------------------------------ 30 3. 發酵豆奶------------------------------------------------------------ 30 4. 豆奶及發酵豆奶甲醇萃取物之製備--------------------------- 31 二、培養基製備---------------------------------------------------------- 31 1. 微生物培養基------------------------------------------------------ 31 2. 細胞培養基--------------------------------------------------------- 31 三、細胞培養------------------------------------------------------------- 31 四、細胞存活率試驗 - MTT assay----------------------------------- 32 五、 NO 含量測定- Griess reaction---------------------------------- 32 六、 iNOS 酵素活性測定--------------------------------------------- 33 七、 iNOS及COX-2蛋白質表現- Western blotting-------------- 33 八、細胞激素分析------------------------------------------------------- 35 （四）統計分析方法---------------------------------------------------------- 36 伍、結果與討論 ---------------------------------------------------------------- 37 （一）實驗結果---------------------------------------------------------------- 37 一、細胞存活率試驗 - MTT assay---------------------------------- 37 二、豆奶及發酵豆奶甲醇萃取物對 LPS 誘導巨噬細胞分泌NO能力之影響---------------------------------------------------- 37 三、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞 iNOS 酵素活性之影響----------------------------------------------------- 42 四、不同劑量 LPS 誘發 RAW 264.7 細胞 iNOS 蛋白質表現之劑量反應----------------------------------------------------------- 42 五、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞 iNOS 蛋白質表現量之影響--------------------------------------------- 43 六、不同劑量 LPS 誘發 RAW 264.7 細胞 COX-2蛋白質表現之劑量反應------------------------------------------------------ 47 七、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞 COX-2 蛋白質表現量之影響--------------------------------------------- 47 八、豆奶及發酵豆奶甲醇萃取物對促發炎細胞激素 (TNF-α、IL-6及IL-1β) 之影響-------------------------------------------- 47 1. 豆奶及發酵豆奶甲醇萃取物對 LPS 誘導 RAW 264.7 細胞生成TNF-α之影響----------------------------------------- 48 2. 豆奶及發酵豆奶甲醇萃取物對 LPS 誘導 RAW 264.7 細胞生成IL-6 之影響------------------------------------------- 48 3. 豆奶及發酵豆奶甲醇萃取物對 LPS 誘導 RAW 264.7 細胞生成IL-1β之影響------------------------------------------- 51 （二）討論---------------------------------------------------------------------- 55 陸、結論 -------------------------------------------------------------------------- 60 柒、參考文獻 -------------------------------------------------------------------- 62 圖次 圖一、大豆中主要異黃酮素之結構-------------------------------------- 5 圖二、革蘭氏陰性菌外膜脂多醣體之化學結構------------------------- 16 圖三、NOS 與NO 之功能------------------------------------------------- 18 圖四、一氧化氮合成路徑--------------------------------------------------- 20 圖五、花生四烯酸代謝路徑------------------------------------------------ 22 圖六、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞存活率之影響-------------------------------------------------------------------- 38 圖七、含 LPS (1μg/ml) 之培養基對 RAW 264.7 細胞存活率之影響----------------------------------------------------------------------- 39 圖八、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞生成 NO 之影響----------------------------------------------------------------- 41 圖九、不同劑量 LPS 誘發巨噬細胞 RAW 264.7 iNOS 蛋白質表現之效應-------------------------------------------------------------- 45 圖十、豆奶及發酵豆奶甲醇萃取物對 LPS誘發 RAW 264.7 細胞 iNOS 蛋白質表現之影響------------------------------------------ 46 圖十一、不同劑量 LPS 誘發巨噬細胞 RAW 264.7 COX-2 蛋白質表現之效應----------------------------------------------------- 49 圖十二、豆奶及發酵豆奶甲醇萃取物對 LPS誘發 RAW 264.7 細胞 COX-2 蛋白質表現之影響-------------------------------- 50 圖十三、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞生成 TNF-α 之影響----------------------------------------------------- 52 圖十四、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞生成 IL-6 之影響-------------------------------------------------------- 53 圖十五、豆奶及發酵豆奶甲醇萃取物對 RAW 264.7 細胞生成 IL-1β 之影響------------------------------------------------------ 54 表次 表一、比較牛奶與豆奶中的營養成分------------------------------------ 9 表二、豆奶及發酵豆奶甲醇萃取物對 LPS 誘發 RAW 264.7 細胞 產生NO的濃度來推測iNOS 酵素活性之影響---------------- 44 | |
| dc.language.iso | zh-TW | |
| dc.title | 利用巨噬細胞株模式探討豆奶與發酵豆奶對發炎反應之影響 | zh_TW |
| dc.title | Effects of soymilk and fermented soymilk on LPS-induced inflammation in RAW 264.7 macrophages | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 黃惠宇 | |
| dc.contributor.oralexamcommittee | 潘崇良,丘志威,游若? | |
| dc.subject.keyword | 豆奶,發酵豆奶,發炎,巨噬細胞, | zh_TW |
| dc.subject.keyword | soymilk,fermented soymilk,inflammation,RAW264.7 macrophages, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-26 | |
| 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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