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研究生:莊家如
研究生(外文):Chia-ju Chuang
論文名稱:活菌體與熱致死乳酸菌對免疫調節影響之比較
論文名稱(外文):Comparative effect of viable and heat-killed lactic acid bacteria on immunomodulation
指導教授:林美吟林美吟引用關係
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:139
中文關鍵詞:活菌體乳酸菌熱致死乳酸菌IL-12p70IFN-γIL-13黏附性
外文關鍵詞:viable lactic acid bacteriaheat-killed lactic acid bacteriaIL-12p70IFN-γIL-13adhesion
相關次數:
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乳酸菌的益生功能不僅在腸道菌相平衡上,在免疫功效上亦具有調節功能,能夠促進人體健康。根據我們先前之研究結果顯示熱致死的乳酸菌具有免疫調節功效,而熱致死的乳酸菌熱比起活菌型態的乳酸菌有著容易儲藏、產品保存期限較長及運輸便利等優點。在本實驗中利用四種不同加熱條件來熱致死11株乳酸菌,接著將熱致死菌及活菌與人體周邊血液單核球細胞共同培養,並以IFN-γ、IL-12p70作為乳酸菌免疫調節功能的指標。另一方面,以Caco-2細胞株為人類腸道上皮細胞膜式,來評估四個熱致死條件及活菌狀態下對其黏附力的影響。綜合IFN-γ、IL-12p70及黏附性試驗結果,選擇三株菌來測其降低人體周邊血液單核球細胞產生IL-13的量,以應證其免疫調節功能。實驗結果顯示出L. salivarius AP-32、E. faecalis YM-73熱致死後比起活菌反而能激發人體周邊血液單核球細胞分泌更高IFN-γ、IL-12p70的量。因此,熱致死乳酸菌具有改善Th1/Th2之間的平衡功能且熱致死的條件於未來乳酸菌製程上也能有所發展及應用。
Probiotic lactic acid bacteria have health-promoting effects not only on balance of intestinal microbiota but also on the immunostimulation. According to our previous study, lactic acid bacteria are reported to be effective on immunomodulation after being heat killed. Heat-killed lactic acid bacteria have the advantages of allowing a longer product shelf-life, easier storage, and transportation. In this study, the effect of 11 strains of heat-killed and viable lactic acid bacteria on the in vitro production of interleukin (IL)-12p70 and interferon (IFN) –γ by human peripheral blood mononuclear cells (hPBMCs) were compared. Furthermore, we also investigated the adhesion ability of 11 strains lactic acid bacteria onto the Caco-2 cells. Based on the results of IL-12p70, IFN-γ production and adhesion ability, we selected three species lactic acid bacteria to measure IL-13 from hPBMCs. The results showed that L. salivarius AP-32, E. faecalis YM-73 heat-killed lactic acid bacteria secreted higher-levels of IL-12p70 and IFN-γ than viable bacteria did in hPBMCs. Our results indicated that the heat-killed lactic acid bacteria also improved Th1/Th2 balance by enhancing IL-12p70, IFN-γ and inhibiting IL-13 production from hPBMCs. Thus, the heat-killed treatment can be used for the industrial preparation of probiotics with the activity of immunomodulation.
中文摘要...............i
Abstract ..............ii
目次..................iii
表目次.................vi
圖目次.................vii
第一章、前言 ...........1
第二章、文獻整理 ....... 3
一、乳酸菌的簡介 ......3
二、乳酸菌的益生功效 ....3
(一) 減緩乳糖不耐症 .....4
(二) 維持腸道菌相平衡 ...5
(三) 預防癌症,抑制腫瘤...5
(四) 降低血清中膽固醇含量 .6
(五) 降血壓 ..............7
(六) 抗發炎能力 .........8
(七) 預防及治療動脈粥狀硬化症..8
三、免疫系統之簡介 ........8
(一)先天性免疫 ..........9
(二)適應性免疫 ........10
四、Th1與Th2細胞之簡介 ...12
(一)免疫細胞與細胞激素 ..12
(二)Th1與Th2免疫細胞 14
五、腸道黏膜免疫系統 ....15
(一) 腸道黏膜之免疫組織 ..15
(二) 腸道相關淋巴組織.....16
(三) 腸道內的免疫反應 .... 16
六、腸道乳酸菌菌相及其作用..18
(一) 乳酸菌菌相及作用 ......18
(二) 乳酸菌對腸道細胞附著性試驗....19
七、乳酸菌與免疫調控相關之文獻 ....20
(一) 衛生學說......................20
(二) 乳酸菌調節機制................21
八、乳酸菌熱致死處理...............23
(一) 乳酸菌熱致死條件 ............23
(二) 食品工廠乳酸菌的製備 .........23
九、場發射掃描電子顯微鏡...........24
(一) 場發射掃描電子顯微鏡介紹......24
(二) 樣品之製作....................25
第三章、實驗目的 ..................32
一、實驗目的 ......................32
二、實驗架構 .....................33
第四章、材料與方法 ...............34
一、實驗材料 ....................34
(一) 試驗菌株 ....................34
(二) 實驗細胞 ...................34
(三) 細胞株 .......................34
(四) 實驗藥品 ..................34
(五) 實驗設備 ...................36
(六) 基本培養基及緩衝溶液之配製 ..36
二、實驗方法 ....................38
(一) 乳酸菌樣品的製備 ..........38
(二) 細胞培養 ...................41
(三) 細胞處理 ...................42
(四) 場發式掃描顯微鏡下觀察乳酸菌..43
(五) 細胞激素測定.................44
(六) Caco-2腸道黏附性試驗..........47
(七) 統計分析 .....................49
第五章、結果與討論 .................50
一、結果 ...........................50
(一) 乳酸菌於不同溫度熱致死結果 .....50
(二) 不同熱致死條件處理的乳酸菌對人體周邊血液單核球(hPBMCs)
產生細胞激素IFN-γ的影響 ......50
(三) 不同熱致死條件處理的乳酸菌對人體周邊血液單核球(hPBMCs)
產生細胞激素IL-12p70的影響.....58
(四) 乳酸菌對腸道細胞株吸附性試驗...65
(五) 場發射掃描電子顯微鏡觀察 .....65
(六) 不同熱致死條件處理的乳酸菌對人體周邊血液單核球(hPBMCs)
產生細胞激素IL-13的影響 ......69
二、討論 ...........................71
第六章、參考文獻 ..................110
附錄 ............................122
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