臺灣博碩士論文加值系統

過敏疾病的產生，簡單而言就是人體的免疫功能失調、出現了不平衡的狀況，引起的一連串不適反應。現今環境及飲食習慣的改變，造成過敏疾病日漸增加。文獻曾指出乳酸菌具有降低過敏症狀的功效，主要機制為提高 T helper 1 (Th1) 免疫反應，抑制 T helper 2 (Th2) 免疫反應，使過敏反應降低。本研究目的利用細胞及動物實驗評估複合乳酸菌產品減低過敏免疫反應的能力，本實驗細胞試驗將乳酸菌產品刺激人類周邊血液單核球細胞 (human periphenal blood mononuclear cells, hPBMC)，以酵素免疫連結吸附分析法 (Enzyme-linked immunosorbent assay, ELISA) 檢測其細胞激素 Interferon-γ (IFN-γ)、Interleukin-4 (IL-4)。動物實驗採用 BALB/c 品系雄性小鼠以隨機方式分為五組，分別為對照組、Kefir Ⅰ、Kefir Ⅱ、Kefir cell free fraction 及市售乳酸菌產品。對照組灌食無菌水，其他各組以無菌水回溶乳酸菌菌粉，其菌數達 107 CFU/ ml，每週灌食七天，一天一次，每隻餵食0.2 ml。實驗為期四週，實驗開始和第二週給予腹腔注射 Ovalbumin (OVA) 第一次注射劑量為 2µg/ mouse，第二次注射劑量為 6µg/ mouse。實驗開始、第一、三和四週以眼窩採血收集血清，檢測免疫球蛋白 E (immunoglobulin E, IgE) 及OVA-specific IgE，四週後犧牲採集脾臟細胞，檢測細胞激素 IFN-γ、IL-12、IL-4 和 IL-5 之分泌量。結果顯示乳酸菌產品能刺激 hPBMC 產生 IFN-γ，減少 IL-4 的分泌，增加 IFN-γ/ IL-4 的比值。在動物試驗方面乳酸菌產品能降低小鼠血清中抗體 IgE 及 OVA-specific IgE，刺激脾臟細胞分泌細胞激素 IFN-γ 和 IL-12，增加 IFN-γ/ IL-4 的比值，使脾臟細胞免疫反應偏向 Th1 反應。綜合本實驗結果三種乳酸菌產品具有降低過敏免疫反應的能力，尤其以 Kefir Ⅱ 效果最佳。

Allergy is a disorder of the immune system. Allergic reactions occur to environmental substances known as allergens. Common allergic diseases include atopic dermatitis, asthma eczema, allergic rhinitis, allergic conjunctivitis and food allergies are the T- helper 1(Th1)/ T-helper 2(Th2) imbalance. There is growing interest in probiotics, not only for treatment of Th1-mediated diseases but also for Th2-mediated diseases. The aim of this study was to evaluate the immunomodulatory effect of probiotic products in vitro and in vivo. Probiotics products included Kefir Ⅰ, Kefir Ⅱ, commercial product group and Kefir cell free fraction. We determine the production of interferon gamma (IFN-γ) and Interleukin-4 (IL-4) cytokines by four different probiotics products in human periphera blood mononuclear cells (hPBMC) model. In animal model, BALB/c mice were radonmly divided into control group, Kefir Ⅰgroup, Kefir Ⅱgroup, Kefir cell free fraction group, commercial product group. The experimental period were 4 weeks, mice were injected with 2 µg/ mouse and 6 µg/ mouse ovalbumin (OVA) mixed with complete Freund’s adjuvant (CFA) at 0 and 2 week. After sacriceing, we measured of serum immunoglobulin E (IgE) and OVA-specific IgE levels and observed the secretion of cytokines IL-4, IL-5, IL-12 and IFN-γby splenocytes. In results, probiotic products induced IFN-γ suppressed IL-4, and increase significantly IFN-γ/ IL-4 (Th1/ Th2) ratio by hPBMC. Probiotic products also decreased significantly the serum IgE and OVA-specific IgE levels of mice, and increased the secretion of cytokines IL-12, IFN-γ and the secretion ratio of INF-γ/ IL-4 by splenocytes. In conclusions, the three probiotic products have immunomodulative effects of decreasing allergy, Kefir Ⅱhas the strongest activity especially.

中文摘要 Ⅰ
英文摘要 Ⅱ
目錄 Ⅲ
表目錄 Ⅵ
圖目錄 Ⅶ

第一章 前言 1

第二章 文獻整理 4
第一節 、免疫與過敏的關係 4
壹 、免疫系統的簡介 4
貳 、腸道免疫系統 5
參 、細胞激素的介紹 5
肆 、分離周邊血液單核球介紹 7
伍 、過敏疾病的介紹 8
陸 、過敏疾病的致病機轉 9
第二節 乳酸菌與過敏的關係 10
壹 、乳酸菌的簡介 10
貳 、乳酸菌的益生特性 11
參 、乳酸菌在免疫方面的功能 11

第三章 材料與方法 20
第一節 實驗材料 20
壹 、試驗樣品 20
貳 、實驗細胞 21
參 、實驗動物 21
肆 、藥品 21
伍 、儀器 23
第二節 細胞試驗 24
壹 、人類周邊血液淋巴球之製備 24
貳 、人類周邊血液淋巴球細胞之計數 24
參 、乳酸菌產品對人類周邊血液單核細胞之免疫激活試驗 25
肆 、酵素免疫連結吸附分析法測定細胞激素 25
第三節 、動物實驗 26
壹 、生長狀況評估 28
貳 、臟器組織相對重量之變化 28
參 、血液樣品收集 28
肆 、血清中 IgE 之測定 28
伍 、血清中 OVA-specific IgE 之測定 29
陸 、脾臟細胞之製備 29
柒 、脾臟細胞之培養及刺激作用 30
捌 、檢測脾臟細胞分泌細胞激素 IFN-γ、IL-4、IL-5 和 IL-12(p40) 30
第四節 統計分析 31

第四章 結果與討論 32
第一節 細胞試驗 32
壹 、乳酸菌產品對人類周邊血液單核球細胞分泌 IFN-γ 和 IL-4 之影響 32
貳 、乳酸菌產品對人類周邊血液單核球細胞分泌 IFN-γ/ IL-4 比值之影響 33
第二節 動物實驗 34
壹 、餵食乳酸菌對 BALB/c 雄鼠每日平均攝食量與體重變化之影響 34
貳 、餵食乳酸菌對 BALB/c 雄鼠臟器組織相對重量之變化 34
參 、餵食乳酸菌對 BALB/c 雄鼠血清中 IgE 抗體及 OVA-specific IgE 抗體濃度之影響 34
肆 、餵食乳酸菌對 OVA 特異性 BALB/c 雄鼠脾臟細胞分泌細胞激素之影響 35
伍 、餵食乳酸菌產品對 OVA 特異性 BALB/c 雄鼠脾臟細胞分泌 IFN-γ/ IL-4 比值影響 37

第五章 結論 39

第六章 參考文獻 68

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