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研究生: 張雅琪
Chang, Ya-Chi
論文名稱: 羅望子木葡聚醣對TNBS誘導小鼠潰瘍性結腸炎的影響
The Effects of Tamarind Xyloglucan on TNBS-induced Ulcerative Colitis in Mice
指導教授: 劉明毅
Liu, Ming-Yie
學位類別: 碩士
Master
系所名稱: 醫學院 - 環境醫學研究所
Department of Environmental and Occupational Health
論文出版年: 2021
畢業學年度: 109
語文別: 中文
論文頁數: 64
中文關鍵詞: 羅望子木葡聚醣 、潰瘍性結腸炎 、促炎細胞因子 、發炎反應 、氧化壓力
外文關鍵詞: Tamarind xyloglucan, ulcerative colitis, cytokines, inflammation, oxidative stress
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  • 潰瘍性結腸炎是發生於結腸部位的慢性炎症,屬於發炎性腸道疾病的一種,常見的症狀為腹痛、腹瀉、胃腸道出血,嚴重則會導致罹患大腸癌的風險增加。然而目前對於此疾病的發病原因還尚未確定,遺傳易感性、環境因素及免疫系統失調皆可能與潰瘍性結腸炎的發展有關。羅望子木葡聚醣(TXG)是一種來自於羅望子樹種子中的天然多醣類,在過去的研究中發現其具有抗發炎及抗氧化的功能,能夠預防葡聚糖硫酸鈉(DSS)所誘導的慢性結腸炎的發生,然而目前還沒有將羅望子木葡聚醣應用在三硝基苯磺酸(TNBS)所誘導的急性結腸炎治療的相關研究。因此,本研究的目的為探討使用羅望子木葡聚醣對於治療TNBS所誘導的小鼠急性結腸炎的潛在應用價值。將小鼠以直腸給藥方式打入TNBS進行結腸炎的誘導後,以餵管方式每天給予一次羅望子木葡聚醣進行治療,並於誘導後第三天與第五天進行犧牲。實驗結束後,採集小鼠血液與結腸進行後續分析,包括血液細胞分析、氧化壓力、發炎反應與結腸組織病理切片。實驗結果顯示,羅望子木葡聚醣能夠顯著減輕小鼠結腸的損傷,並降低COX-2的表現量和促炎細胞因子IL-6、IL-1β以及TNF-α的含量,因此推斷羅望子木葡聚醣可藉由降低發炎反應來減緩TNBS所誘導的小鼠潰瘍性結腸炎。

    Ulcerative Colitis (UC) is a chronic inflammatory disease that occurs in the colon. It is a type of Inflammatory Bowel Disease (IBD) which may lead to abdominal pain, diarrhea, gastrointestinal bleeding, and colorectal cancer. However, the causes of UC have not been identified. It has been suggested that genetic susceptibility, environmental factors and exaggerated immune response may be related to the development of ulcerative colitis. Tamarind xyloglucan (TXG) is a natural polysaccharide isolated from seed of Tamarindus indica which have demonstrated anti-inflammatory and antioxidant capabilities. It can prevent the chronic colitis induced by dextran sodium sulfate (DSS). Therefore, the purpose of this study was to investigate whether tamarind xyloglucan can be used to treat TNBS-induced colitis in mice by reducing oxidative stress and inflammatory response. Colitis was induced by intrarectal administration of a single dose of 2,4,6-trinitrobenzene sulfonic acid (TNBS). After TNBS administration, mice were treated with a single daily oral gavage of TXG and were sacrificed on day 3 or day 5. Complete blood count (CBC), oxidative stress, inflammation and histological parameters were analyzed. The results showed that TXG can reduce the expression of COX-2 and the levels of pro-inflammatory cytokines such as IL-6, IL-1β and TNF-α. Therefore, it is concluded that TXG may attenuate ulcerative colitis by reducing inflammation.

    摘要 I Abstract II 誌謝 VI 目錄 VIII 表目錄 X 圖目錄 XI 壹、研究背景 1 1.1 發炎性腸道疾病(Inflammatory bowel disease, IBD) 1 1.2 潰瘍性結腸炎(Ulcerative colitis, UC) 2 1.3 潰瘍性結腸炎之發病機制 3 1.3.1 過敏反應 4 1.3.2 氧化壓力 4 1.3.3 發炎反應 5 1.4 潰瘍性結腸炎之治療 6 1.5 羅望子木葡聚醣(Tamarind xyloglucan, TXG) 7 貳、研究目的 9 參、材料與方法 10 3.1 實驗材料 10 3.1.1 實驗動物 10 3.1.2 實驗器材 10 3.1.2.1 實驗藥品 10 3.1.2.2 抗體 12 3.1.2.3 實驗試劑組 12 3.1.2.4 實驗耗材 13 3.1.2.5 實驗儀器 14 3.1.2.6 分析軟體 15 3.2 實驗方法 15 3.2.1 實驗動物處理 15 3.2.2 實驗藥劑配置 16 3.3 實驗分析 16 3.3.1 臨床表現之評估 16 3.3.2 樣本採集 17 3.3.3 血液樣本採集 17 3.3.4 結腸組織切片 17 3.3.5 樣本中蛋白質含量測定 17 3.3.6 全血球細胞計數 18 3.3.7 測定結腸中一氧化氮(NO)的濃度 18 3.3.8 測定結腸脂質過氧化(Lipid peroxidation)的程度 18 3.3.9 測定結腸組織中發炎相關細胞因子之含量 18 3.3.10 測定小鼠體內組織胺(Histamine)的濃度 19 3.3.11 測定小鼠體內免疫球蛋白E(IgE)的濃度 19 3.3.12 西方墨點分析法(Western blot) 19 3.4 統計分析 20 肆、實驗結果 21 4.1 羅望子木葡聚醣對結腸炎小鼠體重之影響 21 4.2 羅望子木葡聚醣對結腸炎小鼠疾病活動指數(DAI)之影響 21 4.3 羅望子木葡聚醣對結腸炎小鼠攝食量之影響 21 4.4 羅望子木葡聚醣對結腸炎小鼠結腸長度之影響 21 4.5 羅望子木葡聚醣對結腸炎小鼠結腸水腫程度之影響 22 4.6 羅望子木葡聚醣對結腸炎小鼠脾臟水腫程度之影響 22 4.7 羅望子木葡聚醣對結腸炎小鼠結腸組織型態之影響 22 4.8 羅望子木葡聚醣對結腸炎小鼠結腸黏蛋白之影響 23 4.9 羅望子木葡聚醣對結腸炎小鼠氧化壓力之影響 23 4.10 羅望子木葡聚醣對結腸炎小鼠血球之影響 23 4.11 羅望子木葡聚醣對結腸炎小鼠過敏反應之影響 23 4.12 羅望子木葡聚醣對結腸炎小鼠發炎反應之影響 24 4.13 羅望子木葡聚醣對結腸炎小鼠發炎相關細胞因子之影響 24 伍、研究討論 26 陸、結論 33 柒、參考文獻 34 捌、圖表 50 玖、附錄 63

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