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研究生:陳優俐
研究生(外文):Yu-Li Chen
論文名稱:利用DNBS誘發產生發炎性腸炎之豬隻動物模式開發
論文名稱(外文):The development of DNBS induced porcine inflammatory bowel disease model.
指導教授:陳有任
指導教授(外文):Yu-Jen Chen
學位類別:碩士
校院名稱:元培科技大學
系所名稱:醫學檢驗生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
畢業學年度:102
語文別:中文
論文頁數:95
中文關鍵詞:發炎性腸炎雙水球誘發導管內視鏡24 -二硝基苯磺酸
外文關鍵詞:Inflammatory bowel diseasePig (Porcine)double-balloon catheterendoscopic24- Dinitrobenzene sulphonic acid(DNBS)
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發炎性腸炎(Inflammatory bowel disease;IBD),主要包括克隆氏症(Crohn's disease;CD)與潰瘍性結腸炎(Ulcerative colitis;UC)。這兩者均為慢性腸道炎症疾病,其致病原因具多樣性,包括環境、細菌感染、飲食習慣、基因及免疫系統等各方面,但至今仍未能證實其確切發病機轉。以藥物控制此病仍是目前主要的治療方式,但臨床上藥物的選擇有限,開發新藥勢在必行,而合適的動物模式更是開發新藥中重要的一環。本研究選擇消化系統及生理結構與人類最為相仿的肉豬作為試驗動物,使用2,4- Dinitrobenzene sulphonic acid (DNBS),以自製之雙水球誘發導管將DNBS置於腸道內,利用化學藥物刺激腸道誘發產生類似發炎性腸炎之病變,再以內視鏡觀察不同誘發條件之腸道病變,建立發炎性腸炎之動物模式。最佳誘發條件為DNBS 150 mg/mL、體積40 mL,誘發刺激1小時,並於刺激後以100 mL蒸餾水清洗腸道殘留DNBS。於最佳條件下誘發豬隻產生腸道病變後,以市售藥品(Pentasa)與安慰劑(starch)治療。6隻試驗豬隻各別連續餵食市售藥品或安慰劑 28天,以內視鏡觀察腸道病理變化,兩者間具有顯著差異,表示市售藥品可有效緩解豬隻經DNBS誘發產生之腸道炎症反應。本研究成功建立出發炎性腸炎豬隻動物誘發模式,並確認可使用於評估發炎性腸炎藥物。
Inflammatory bowel disease (IBD) is including Crohn's disease (CD) and ulcerative colitis (UC). Both of which are chronic inflammatory bowel disease, its causes is not yet clear. Patients with inflammatory bowel disease currently only symptom relief by tracing drugs.in severe cases, surgical excision of the lesion are required, but there is still the risk of recurrence. In this study, we used pigs as test animals, because their digestive system and physical structure are most similar to human. Weused double-balloon catheter to induce of 2,4 - Dinitrobenzene sulphonic acid (DNBS), is one kind of chemical drug with low toxicity and suitable for medium to large animals, for mimic stimulating the similar symptom of IBD. We use different conditions to induce intestinal inflammation and view the wound by endoscopic. We found the best induce condition is: DNBS 150 mg / mL,40 mL,induce for 1 hour, and wash out intestinal drug residues by 100mL distilled water. Thereafter, we investigated the effects of commercial drug (Pentasa) and starch. At first, we used the best induced condition to create intestinal inflammation, and then, pigs are taking drug or starch for 28 days. By using wound injury total score and average rate of wound healing,the commercial drug decreased the wound inducing by DNBS, and starch had no effect for the wound. There are significantly different results between these two treatment.Thus, this study successfully established an induced-animal mode of inflammatory bowel disease in pigs. In the future, the development of new drugs for inflammatory bowel disease, can based on this animal mode for testing the drug efficacy to accelerate the drug into clinical trials. Helping more patients with inflammatory bowel disease have more therapy options.
謝誌 I
中文摘要 II
英文摘要 III
目錄 IV
圖目錄 VI
表目錄 VII
附錄圖表目錄 VIII
中英文名詞縮寫對照表 IX
第一章緒論 1
1.1 發炎性腸炎(Inflammatory bowel disease;IBD) 1
1.2 克隆氏症 (Crohn's disease;CD) 2
1.3 潰瘍性結腸炎 (Ulcerative colitis;UC) 2
1.4 發炎性腸炎臨床診斷方法 3
1.5 發炎性腸炎臨床治療說明 6
1.6 發炎性腸炎與結直腸癌 9
1.7 發炎性腸炎誘導模式 9
1.8 發炎性腸炎誘導動物模式 11
第二章材料方法 15
2.1 試驗材料 15
2.1.1 試驗藥物 15
2.1.2 試驗儀器 15
2.1.3 試驗耗材 16
2.2 試驗方法 17
2.2.1 試驗動物飼養 17
2.2.2 誘發藥物配置 17
2.2.3 試驗動物麻醉 17
2.2.4 試驗動物清腸 18
2.2.5 內視鏡臨床觀察 18
2.2.6 誘發產生發炎性腸炎 18
2.2.7動物餵食藥物方法 19
2.2.8結腸傷口損傷程度 19
2.2.9動物腸道生檢取樣 21
2.2.10組織切片 21
2.2.11切片染色及判讀 21
2.2.12統計方法 22
2.3 試驗設計 23
2.3.1 建立DNBS刺激誘發豬隻IBD模式 23
2.3.2 使用最佳IBD誘發條件進行安慰劑測試 27
2.3.3 評估市售藥品於IBD豬隻動物模式之療效 29
第三章結果 31
3.1 建立DNBS刺激誘發豬隻IBD模式 31
3.1.1 條件一 31
3.1.2 條件二 34
3.1.3 條件三 38
3.1.4 條件四 42
3.2 使用最佳IBD誘發條件進行安慰劑測試 47
3.3 評估市售藥品於IBD豬隻動物模式之療效 53
第四章討論 61
參考文獻 66
附錄 76
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