不同品種乳酸菌對於預防沙門氏桿菌脂多醣體所造成上皮細胞屏障功能損害之體外試驗模型研究

In-vitro Prevention of Salmonella Lipopolysaccharide-induced Damages in Epithelial Barrier Function by Various Lactobacillus Strains

楊俊仁(Chun-Yan Yeung)

指導教授 ： 劉宣良

國立臺北科技大學/工程學院/工程科技研究所/博士(2013年)

https://doi.org/10.6841/NTUT.2013.00732

摘要

研究顯示乳酸菌對沙門氏桿菌感染有很好的抗發炎效果，其作用為維持腸道上皮細胞密合區的完整性，在避免細菌的侵入有其重要性。本研究旨在探討在已發炎的腸道上皮細胞上，乳酸菌是否可有效調控腸道上皮細胞密合區蛋白的表現和分佈。使用轉移盤共同培養模型，沙門氏桿菌脂多醣體添加於已極化之Caco-2細胞的上層，周邊血液單核球細胞則共同培養於下層。被脂多醣體刺激活化的Caco-2細胞則與不同品種的乳酸菌共同培養。密合區的完整性可經由測量Caco-2細胞層的電組來判斷。而密合區蛋白(閉鎖小帶ZO-1)的表現及分佈可使用西式墨點法及免疫螢光來確認。不同品種的乳酸菌對於Caco-2細胞層的完整性有不同的調控能力，而脂多醣體被証實可用於破壞腸道上皮細胞的屏障及改變閉鎖小帶(ZO-1)的分佈。我們的數據顯示乳酸菌可以弱化因使用沙門氏桿菌脂多醣體導致的腸道上皮細胞屏障的破壞程度，同樣也顯示乳酸菌的品種與維持密合區完整性和表現有關。我們的研究結果顯示活的益生菌可以改善腸道上皮細胞的屏障，且這可解釋益生菌在臨床上對於活體效益的潛在機制。

關鍵字

乳酸菌 ； 沙門氏桿菌 ； 脂多醣體 ； 密合區 ； Caco-2細胞

並列摘要

Lactobacillus shows beneficial anti-inflammatory effects to Salmonella infection. The maintenance of the tight junction (TJ) integrity plays an importance role in avoiding bacterial invasion. Whether Lactobacillus could be used to regulate the TJ protein expression and distribution in inflamed intestinal epithelial cells was determined. Using the transwell co-culture model, Salmonella lipopolysaccharide (LPS) was apically added to polarized Caco-2 cells co-cultured with peripheral blood mononuclear cells in the basolateral compartment. LPS-stimulated Caco-2 cells were incubated with various Lactobacillus strains. TJ integrity was determined by measuring trans-epithelial electrical resistance across Caco-2 monolayer. Expression and localization of TJ proteins (zonula occludens (ZO)-1) were determined by western blot and immunofluorescence microscopy. Various strains of Lactobacillus were responsible for the different modulations of cell layer integrity. LPS was specifically able to disrupt epithelial barrier and change the location of ZO-1. Our data demonstrate that Lactobacillus could attenuate the barrier disruption of intestinal epithelial cells caused by Salmonella LPS administration. We showed that Lactobacillus strains are associated with the maintenance of the tight junction integrity and appearance. In this study we provide insight that live probiotics could improve epithelial barrier properties and this may explain the potential mechanism behind to their beneficial effect in-vivo.

並列關鍵字

Lactobacillus ； Salmonella ； lipopolysaccharide ； tight junction ； Caco-2 cells

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