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研究生:廖昱婷
研究生(外文):Yu-Ting Liao
論文名稱:乳酸菌之免疫調節作用
論文名稱(外文):Immunomodulatory activity of lactic acid bacteria
指導教授:林美吟林美吟引用關係
指導教授(外文):Meei-Yn Lin
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:乳酸菌免疫調節巨噬細胞
外文關鍵詞:Lactic acid bacteriaImmunomodulatoryMacrophage
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在本實驗中,我們測試8株乳酸菌對小鼠巨噬細胞株J774A.1的免疫刺激的影響,8株乳酸菌包括:L. acidophilus 4356、L. acidophilus LA-1、L. bulgaricus 448、L. bulgaricus 449、B. longum B6、B. longum 15708、S. thermophilus MC及S. thermophilus 573。乳酸菌樣品包括乳酸菌活菌株、熱致死菌、胞內液、胞外液、粗細胞壁萃取物、染色體DNA及乳酸菌發酵乳的乳清。測試的項目包括:刺激巨噬細胞株J774A.1的增生、激活細胞株分泌TNF-α及IL-6的量、刺激巨噬細胞株J774A.1的吞噬能力。結果顯示,乳酸菌株對細胞株J774A.1增生的調節是經由107 cfu/ml活菌株、0.05 μg/ml Genomic DNA及107 cfu/ml菌數之發酵乳乳清的刺激,但刺激程度會依菌株及菌數而有所不同。8株乳酸菌在1,000及100 μg/ml細胞壁粗萃物,或是106-108 cfu/ml菌數之發酵乳乳清與巨噬細胞株共培養,具有顯著刺激巨噬細胞株分泌TNF-α的能力。只有1,000、100 μg/ml細胞壁粗萃物與部分活菌樣品,如107 cfu/ml之L. acidophilus LA-1;107-108 cfu/ml之L. bulgaricus 448;106-107 cfu/ml之B. longum 15708及S. thermophilus MC,可以刺激巨噬細胞株分泌IL-6。乳酸菌的熱致死菌體、胞內液、胞外液、Genomic DNA及發酵乳乳清都可以刺激巨噬細胞株增加吞噬的能力。實驗結果中顯示,乳酸菌調節免疫能力的程度會因菌株和菌數不同,顯示了菌株依賴性(Strain-dependent)。本實驗結果亦顯示,乳酸菌可以激活巨噬細胞,但不至於產生發炎反應,故可以增強免疫系統的能力。
The stimulatory effect of eight strains of lactic acid bacteria including L. acidophilus 4356, L. acidophilus LA-1, L. bulgaricus 448, L. bulgaricus 449, B. longum B6, B. longum 15708, S. thermophilus MC and S. thermophilus 573 on murine macrophage J774A.1 cell line were examined. The fractions of lactic acid bacteria including viable bacteria, heat-killed bacteria, intracellular extract, extracellular extract, crude cell wall, chromosome DNA, and whey of fermented milk were used for this study. The testing items include proliferation, the secretion of TNF-α and IL-6, and phagocytosis of murine macrophage J774A.1 cell line. The result showed that lactic acid bacteria activate the macrophages proliferation by 107 cfu/ml of viable bacteria, fermented milk and 0.05 μg/ml of genomic DNA, respectively. And the degree depended on the difference of strains and numbers of bacterium. Co-culture with 1,000 and 100 μg/ml crude cell wall extract or whey of fermented milk which containing 106-108 cfu/ml lactic acid bacteria with macrophage cell line significantly increased its production of TNF-α. Crude cell wall extracts (1,000 and 100 μg/ml) and viable bacteria (107 cfu/ml L. acidophilus LA-1, 107-108 cfu/ml, L. bulgaricus 448, 106-107 cfu/ml B. longum 15708 and S. thermophilus MC) could stimulate the production of IL-6 by macrophage cell line. Many fractions of lactic acid bacteria could stimulate phagocytosis of macrophage cell line, including heat-killed bacteria, intracellular extract, extracellular extract, genomic DNA and whey of fermented milk. Immunomodulatory activity of lactic acid bacteria depended on difference of strains and numbers of bacterium, and expressed strain-dependent manners. In our results also indicate that lactic acid bacteria can stimulate the macrophage cell line and may improve immune system, but not to such an extent as to inflammation.
目 錄
頁次
中文摘要……………………………………………………………vii
英文摘要……………………………………………………………viii
表目錄………………………………………………………………iv
圖目錄………………………………………………………………v
第一章 前言………………………………………………………1
第二章 文獻整理…………………………………………………3
一、乳酸菌簡介………………………………………………3
二、乳酸菌的功效……………………………………………3
三、腸道免疫…………………………………………………6
1. 免疫反應的簡介 ……………………………………6
2. 腸黏膜之免疫組織 …………………………………10
3. 腸道內的免疫反應 …………………………………12
四、巨噬細胞…………………………………………………14
1. 巨噬細胞的反應與活化 ……………………………14
2. 巨噬細胞的吞噬作用 ………………………………15
3. 巨噬細胞的殺菌和細胞毒殺作用 …………………15
4. 巨噬細胞分泌之物質 ………………………………16
五、乳酸菌與免疫調節作用…………………………………17
第三章 材料與方法………………………………………………25
一、材料………………………………………………………25
1. 試驗菌株 ……………………………………………25
2. 細胞株 ………………………………………………25
3. 實驗藥品 ……………………………………………25
4. 基本培養基與緩衝溶液配製 ………………………27
5. 實驗設備 ……………………………………………29
二、方法 ……………………………………………………31
1. 乳酸菌菌種保存 ……………………………………31
2. 乳酸菌菌株之活化 …………………………………31
3. 乳酸菌菌數之測定 …………………………………31
4. 乳酸菌樣品之製備 …………………………………32
5. J774A.1細胞株之抗凍培養基配製…………………35
6. J774A.1細胞株之保存………………………………35
7. J774A.1細胞株之解凍與培養………………………35
8. 利用MTT色度分析法測試J774A.1細胞株之增生率 36
9. J774A.1吞噬能力分析………………………………37
10. 細胞激素分泌試驗 …………………………………39
11. TNF-α及IL-6之測試…………………………………40
12. 統計分析 ……………………………………………42
第四章 結果………………………………………………………43
一、乳酸菌對巨噬細胞株J774A.1增生之影響 ……………43
二、乳酸菌對巨噬細胞株J774A.1分泌TNF-α之影響………46
三、乳酸菌對巨噬細胞株J774A.1分泌IL-6之影響 ………50
四、乳酸菌對巨噬細胞株J774A.1吞噬能力之影響 ………51
第五章 討論………………………………………………………78
一、乳酸菌對巨噬細胞株J774A.1增生之影響 ……………78
二、乳酸菌對巨噬細胞株J774A.1分泌TNF-α之影響………79
三、乳酸菌對巨噬細胞株J774A.1分泌IL-6之影響 ………82
四、乳酸菌對巨噬細胞株J774A.1吞噬能力之影響 ………83
五、綜合討論………………………………………………………85
第六章 參考文獻…………………………………………………87
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