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研究生:何志軒
論文名稱:乳酸菌抑制脂多醣誘導Caco-2細胞發炎之研究
論文名稱(外文):Inhibitory effects of lactic acid bacteria on lipopolysaccharide induced inflammation in Caco-2 cells
指導教授:魏佳俐魏佳俐引用關係
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生化科技學系研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
畢業學年度:103
語文別:中文
中文關鍵詞:乳酸菌脂多醣Caco-2RAW 264.7
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腸道免疫細胞占人體的總免疫系統高達70〜80%,因此腸道在面對不斷的免疫刺激、耐受性和免疫反應之間的自體平衡,成為腸道免疫系統控制感染源的必要的條件,由微生物病原體所引發的腸道感染往往會誘導許多發炎性病症,包括食物過敏、發炎性腸道疾病(Inflammatory bowel disease;IBD)和大腸癌。
目前已有許多研究指出,乳酸菌(Lactic acid bacteria)能透過改善腸道微生物菌群和調節免疫系統來達到促進腸道健康的效果。因此,本研究以七株Lactobacillus屬及一株Lactococcus屬的乳酸菌為材料。首先以脂多醣(lipopolysaccharide;LPS)誘發RAW 264.7小鼠巨噬細胞株發炎,同時加入熱處理乳酸菌。結果顯示YM0039(Lactobacillus brevis)及YM0512(Lactobacillus plantarum)對NO生成,與已商業化的Shirota(Lactobacillus casei Shirota)和LGG(Lactobacillus rhamnosus GG)的抑制效果相當(49%~55%)。
接著,利用LPS誘發Caco-2人類上皮大腸癌細胞株發炎。在上述八株熱處理乳酸菌與LPS共同處理時,YM0039雖然對促發炎白細胞介素-8(interleukin-8;IL-8)mRNA表現和蛋白質分泌的抑制效果(43%及34%)略低於Shirota(69%及50%)和LGG(74%及43%),然在統計分析上三者均是無顯著差異(p<0.01)。而在乳酸菌處理10小時後,再以LPS誘發Caco-2細胞發炎上,YM0039具有最佳的55% IL-8 mRNA抑制效果,不過仍不及LGG的79%抑制效果。
實驗結果顯示L. brevis YM0039這株乳酸菌具有顯著抗發炎功效,未來希望能將這株乳酸菌當作一種輔助性的健康食品,應用在IBD的症狀減緩和腸道保健。
中文摘要 1
英文摘要(Abstract) 3
壹、 前言 5
一、 腸道上皮細胞的免疫表現 5
二、 發炎性腸疾病 5
三、 引發腸道發炎的致病菌群 7
四、 腸道致病菌所誘導的發炎反應 8
五、 乳酸菌簡介 11
六、 乳酸菌對於IBD患者的療效 13
貳、 研究動機與策略 15
參、 材料與方法 16
一、 實驗材料 16
(一)、化學藥品、材料與酵素 16
(二)、儀器 17
(三)、細胞株與菌株 18
(四)、培養基及化學試劑配法 18
二、 實驗方法 20
(一)、乳酸菌製備 20
(二)、細胞培養 20
(三)、細胞計數 21
(四)、乳酸菌對LPS刺激RAW 264.7細胞株之NO生成量分析 21
(五)、細胞存活率測試 22
(六)、乳酸菌對LPS刺激Caco-2細胞之處理 23
(七)、RT-PCR 23
(八)、ELISA分析 25
(九)、統計分析 26
肆、 結果 27
一、 乳酸菌同步添加對LPS刺激RAW 264.7細胞株之細胞存活及NO生成影響27
二、 乳酸菌同步添加對LPS刺激CACO-2細胞株之IL-8 MRNA表現影響 28
三、 乳酸菌同步添加對LPS刺激CACO-2細胞株之IL-8分泌蛋白表現影響 28
四、 乳酸菌前處理對LPS刺激CACO-2細胞株之IL-8 MRNA表現影響 29
伍、 討論 30
陸、 參考文獻 34
柒、 圖表 39
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