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研究生:林佑軒
研究生(外文):You-Shuan Lin
論文名稱:液態優酪乳對卵白蛋白誘導小鼠之呼吸道過度反應的免疫調節作用
論文名稱(外文):The Immunnomodulatory Effects of Drink Yoghurt on OVA-induced Airway Hyperresponsiveness in Murine Model
指導教授:謝長奇謝長奇引用關係
指導教授(外文):Chang-Chi Hsieh
口試委員:吳勇初詹明修
口試委員(外文):Yun-Chu WuMing-Shiou Jan
口試日期:2015-01-22
學位類別:碩士
校院名稱:東海大學
系所名稱:畜產與生物科技學系
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:100
中文關鍵詞:呼吸道過度反應過敏益生質益生菌優酪乳
外文關鍵詞:Airway hyperresponsivenessAllergyPrebioticsProbioticsYoghurt
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過敏性氣喘是一種慢性的支氣管疾病,伴隨著肺部嗜酸性白血球浸潤肺部、血中免疫球蛋白IgE增加及呼吸道過度反應。研究顯示富含益生菌與益生質的優酪乳可減緩過敏小鼠的發炎症狀、降低IgE及過敏細胞激素IL-4生成,細胞與動物實驗也顯示益生菌能減緩雞卵白蛋白(ovalbumin,OVA)致敏之抗原特異性IgE的含量。本實驗將探含有嗜酸乳桿菌、保加利亞乳桿菌、雙歧乳桿菌、嗜熱乳鏈球菌、腸球菌與果寡糖的東海優酪乳在雞卵白蛋白誘發過敏性氣喘之小鼠的免疫調節作用,藉由分析脾臟、肺部沖洗液、呼吸道周邊淋巴結及腸繫膜淋巴結中淋巴球的表面抗原(CD3、CD4、CD8、CD19、CD25、CD45、CD278、Tim-3)、雞卵白蛋白特異性免疫球蛋白(anti-OVA IgE、anti-OVA IgG1)及細胞激素(IFN-γ、IL-4、IL-5);另一方面測定優酪乳調節過敏原OVA誘導呼吸道阻力及乙醯甲膽鹼(methacholine) 誘導呼吸道過度反應之變化;以蘇木紫與伊紅染色(hematoxylin-eosin stain)、甲苯胺藍染色(toluidine blue)及過碘酸希夫染色(Periodic acid-Schiff stain)評估肺部組織病理之發炎細胞浸潤、肥大細胞脫顆粒現象及杯狀細胞分泌黏液程度。結果顯示,富含益生菌與益生質之優酪乳可顯著減少肺部沖洗液及脾臟淋巴細胞的IL-4、IL-5分泌量及提高IFN-γ分泌量,顯著降低血清及肺部沖洗液中OVA的特異性抗體IgE、IgG1並提高IgG2分泌量,抑制肥大細胞脫顆粒降低肺部發炎反應進而顯著降低呼吸道過度反應及呼吸道阻力。綜上所述,富含益生菌與益生質之優酪乳能透過 T 輔助細胞的免疫調節進而影響 B 細胞的分化,抑制過敏相關免疫球蛋白分泌,減少肥大細胞的脫顆粒作用而降低發炎介質所造成之呼吸道過度反應,而達到改善氣喘的目的。
關鍵詞:呼吸道過度反應、過敏、益生質、益生菌、優酪乳

Allergic asthma is a chronic disease with airway hyperresponsiveness, infiltration of eosinophils into the lungs and increase of serum IgE. Previous studies indicated that the yoghurt could reduce inflammatory effect, suppress allergic cytokine IL-4 and decrease serum IgE in murine model. The in vivo and in vitro studies show probiotics can attenuate the concentration of IgE, ovalbumin (OVA)-specific IgE in OVA sensersized mice. In this study, we administered yoghurt (contain Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium lactis, Streptococcus thermophilus, Enterococcus faecium and fructooligosaccharide) to determine the effect on the OVA-induce allergic asthma in murine model. Lymphocytes were harvested from spleen, bronchoalveolar lavage fluid (BALF), airway-draining and mesenteric lymph nodes. The cluster of differentiation marker (CD3, CD4, CD8, CD19, CD25, CD45, CD278 and Tim-3), OVA-specific immunoglobulin (anti-OVA IgE and anti-OVA IgG1) and cytokines (IFN-γ, IL-4 and IL-5) were determined. The pulmonary pathological sections including H&E, toluidine blue and PAS satin were determined inflammatory cells infiltration, degranulation of mast cell and mucus secretion. Results indicated that yoghurt enriched with probiotics and prebiotics could reduce the concentration of IL-4 and IL-5 but increase IFN-γ in BALF and splenocytes, suppressed degranulation of mast cell and attenuated inflammation in lung. The production of anti-OVA IgE and IgG1 were decrease significantly. Taking these results in conclusion, yoghurt enriched with probiotics and prebiotics can regulated airway hyperresponsiveness via suppressed the allergic immune response including T helper cells polarization, B cell differentiation and further stabilized mast cells degranulation.
Keyword: Airway hyperresponsiveness, Allergy, Prebiotics, Probiotics, Yoghurt

目次
壹、 摘要
貳、 前言
參、 文獻回顧
一、 氣喘之文獻探討
(一) 氣喘定義
(二) 氣喘流行病學
(三) 氣喘病理學
(四) 氣喘機制
(五) 氣喘動物模式
(六) 相關過敏生物分子
1 細胞激素
2 免疫球蛋白
3 嗜酸性白血球
4 肥大細胞
5 杯狀細胞
二、 優酪乳之文獻探討
(一) 優酪乳之定義
(二) 乳酸菌之文獻探討
(三) 益生菌之定義
(四) 益生菌及腸道菌相
(五) 益生菌之功能文獻探討
三、 益生質之文獻探討
(一) 益生質之定義
(二) 益生質之種類來源
(三) 益生質與腸道菌相
(四) 益生質功效
(五) 益生質之食品應用

肆、 研究動機與目的
伍、 材料與方法
一、 實驗方法
(一) 產品檢驗與實驗分組
(二) 呼吸道過度反應之動物模式
(三) 檢體採集與分析
二、 實驗材料
(一) 藥品
(二) 設備、器材
陸、 統計分析
柒、 結果
一、 優酪乳穩定性檢測及小鼠腸道菌相
(一) 優酪乳菌種鑑定
(二) 優酪乳菌數檢測
(三) 小鼠腸道菌相檢測
二、 小鼠呼吸道過度反應之免疫調節
(一) 小鼠體重、脾臟重量之變化
(二) 呼吸道過度反應中浸潤肺部淋巴球細胞組態
(三) 呼吸道過度反應中呼吸道週邊淋巴細胞亞族群之變化
(四) 呼吸道過度反應中脾臟淋巴細胞亞族群之變化
(五) 呼吸道過度反應中腸繫膜淋巴細胞次族群之變化
(六) 呼吸道過度反應中肺部沖洗液細胞激素之變化
(七) 呼吸道過度反應中血清與肺部沖洗液抗體之變化
(八) 呼吸道過度反應之變化
(九) 肺臟病理切片分析
(十) 肺臟免疫組織化學染色分析
三、 優酪乳之生物安全性檢測
(一) 小鼠脾臟免疫細胞增生能力之變化
(二) 小鼠吞噬細胞活性之變化
(三) 小鼠自然殺手細胞活性之變化
捌、 討論
玖、 結論
壹拾、 參考文獻
壹拾壹、 附件

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