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研究生:王瑋琦
研究生(外文):Wei-Kee Wang
論文名稱:乳酸菌之抗發炎與改善潰瘍性腸炎功效評估
論文名稱(外文):Effects of lactic acid bacteria on amelioration of inflammatory response and ulcerative colitis
指導教授:蔡英傑蔡英傑引用關係
指導教授(外文):Ying-Chieh Tsai
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生化暨分子生物研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:74
中文關鍵詞:乳酸菌發炎潰瘍性結腸炎
外文關鍵詞:lactic acid bacteriainflammationulcerative colitis
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發炎為人體免疫系統為了抵抗外來的致病原所產生的一種保護機制。當發炎反應失去控制 (dysregulation) 而持續發生會導致過度發炎,因而引發或輕或重的病理後果 (pathological conditions) 並導致疾病的產生,本論文研究主題潰瘍性結腸炎 (ulcerative colitis; UC) 就是一種腸道持續性發炎的疾病。目前已有許多的研究指出,乳酸菌 (lactic acid bacteria; LAB) 具有免疫調節的能力,因此,我們利用脂多醣 (lipopolysaccharide; LPS) 引發RAW 264.7小鼠巨噬細胞株發炎的細胞模式,及葡聚糖硫酸酯鈉鹽 (dextran sulfate sodium; DSS) 誘發潰瘍性結腸炎疾病產生的動物模式,評估乳酸菌抗發炎及改善UC此發炎性疾病的能力。
首先以LPS誘發RAW 264.7小鼠巨噬細胞株,同時加入熱殺死乳酸菌,以促發炎介質 (pro-inflammatory cytokine),包括一氧化氮 (nitric oxide; NO)、腫瘤壞死因子 (tumor necrosis factor α; TNF-α) 及前列腺素E2 (prostaglandin E2; PGE2)的濃度下降為指標,評估待測乳酸菌的抗發炎能力。從100株待測乳酸菌中篩選出了7株能有效抑制上述促發炎介質產生的乳酸菌,由細胞實驗結果進入潰瘍性結腸炎動物實驗。結果顯示,餵食5048及7036這兩株乳酸菌的實驗小鼠,其潰瘍性結腸炎的症狀,包括:體重減輕、脾臟重量增加及結腸長度縮短的臨床現象,都有明顯改善。同時,其結腸組織中促發炎介質包括:TNF-α、介白質-1β (interleukin-1β; IL-1β) 及介白質-6 (interleukin-6; IL-6) 的濃度也有顯著性的下降。此外,在組織學分析的結果也發現,餵食5048及7036的實驗小鼠可以有效的回復結腸腸壁結構的完整性,同時,骨髓過氧化酶 (myeloperoxidase; MPO) 的活性下降,間接表示結腸組織因發炎產生的嗜中性球浸潤現象也獲得改善。
依據以上實驗結果,顯示5048及7036這兩株乳酸菌,在細胞模式能降低LPS誘發RAW 264.7細胞產生的促發炎介質濃度,包括:NO、TNF-α、PGE2;在DSS誘發小鼠潰瘍性結腸炎的動物模式中,可以改善潰瘍性結腸炎造成小鼠體重減輕、脾臟重量增加及結腸長度縮短等現象。
由本論文實驗結果顯示5048及7036這兩株乳酸菌具有改善發炎疾病的能力,未來我們也希望能將這兩株乳酸菌當作一種輔助性的健康食品,應用在潰瘍性結腸炎患者疾病的減緩上。

Inflammation is a kind of protective response triggered by infection or tissue injury. It is generally thought that a controlled inflammatory response is beneficial, but it can become harmful if dysregulated. Ulcerative colitis (UC) is an example of disease causing by excessive inflammation, however, current treatments for it are relatively ineffective. Up to the present, many researchers have demonstrated that lactic acid bacteria (LAB) have immunomodulating potential. Therefore, we evaluated the effect of LABs isolated from variety of sources on lipopolysaccharide (LPS)-stimulated RAW 264.7 inflammatory cell model and dextran sulfate sodium (DSS)-induced UC murine model.
We screened the inhibitory effect of LABs against pro-inflammatory cytokine production, including nitric oxide (NO), tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), induced by LPS. Seven LABs stand out above the rest, due to they can reduce the release of these cytokines effectively. Further, seven candidate LABs were compared the effect by oral administration on the murine model of UC. Among seven LABs, 5048 and 7036, are the most effective in alleviation of UC symptoms, including decrease of body weight loss, inhibition of spleen weight increase and prevention of colon shortening. The concentration of pro-inflammatory cytokines, TNF-α, interleukin-1β (IL-1β), and interleukin-6 (IL-6) in colon tissue are reduced by 5048 and 7036 treatments, too. In histological analysis, the architecture of colonic wall is restored. Furthermore, myeloperoxidase (MPO) activity, the marker of neutrophil infiltration, is also decrease by oral administration of 5048 and 7036.
According to our experiment, we can concluded that 5048 and 7036 can ameliorate the clinical symptoms in mice with UC induced by DSS. Thus, these two LABs could be expected as an auxiliary agent to UC patients.
中英文名詞縮寫對照表                   i
中文摘要                         ii
英文摘要                         iv
第一章 緒論                       1
 一、發炎反應                      1
 二、發炎性細胞激素 (proinflammatory cytokine)  3
 三、潰瘍性結腸炎 (ulcerative colitis; UC)    7
 四、乳酸菌                       9
第二章 實驗材料與方法                  17
一、實驗材料                      17
二、實驗方法                      19
第三章 實驗結果                     28
 一、細胞模式In vitro                 28
 二、動物模式In vivo 第一階段            30
 三、動物模式In vivo 第二階段            32
第四章 討論                       35
第五章 參考文獻                     40
第六章 圖表                       45

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