益生性乳酸菌對Clostridium difficile在Caco-2細胞單層膜吸附之影響

Effect of probiotic lactic acid bacteria on adhesion of Clostridium difficile to Caco-2 cell line monolayer

陳正祥(Cheng-hsiang Chen)

指導教授 ： 顏聰榮

大同大學/工程學院/生物工程學系所/碩士(2004年)

摘要

許多研究指出Clostridium difficile (C. difficile)會吸附在Caco-2細胞上並分泌毒素傷害Caco-2；如果添加高濃度C. difficile分泌的毒素會引起Caco-2細胞死亡。本研究發現Lactobacillus acidophilus GG(L. GG) 與C. difficile共同培養在MRS broth中，L.GG 會抑制C . difficile生長，第16 小時C .difficile生長變慢，至第36 小時生菌數下降。另一項實驗發現Caco-2單層膜在感染C. difficile後，TEER值會下降顯示膜的完整性被破壞。而同時添加C. difficile和益生性乳酸菌L. GG，則TEER值比單獨感染C. difficile高，說明益生性乳酸菌L. GG有防止C. difficile對膜之傷害。結果顯示C. difficile會傷害Caco-2單層膜，而益生性乳酸菌L. GG會抑制C. difficile對Caco-2細胞單層膜傷害，因而添加益生性乳酸菌L. GG有保護腸管道避免C. difficile所引起的偽膜性腸炎。而實驗進一步確認益生性乳酸菌L. GG對C. difficile偒害Caco-2單層膜之保護機制。在顯微鏡下觀察在單層膜上L. GG對C. difficile作用， 可發現L. GG都會纏繞在C. difficile周圍，如果C. difficile是吸附在單層膜tight junction處L. GG同樣也會吸附在附近位置。

關鍵字

Caco-2細胞 ； 益生性乳酸菌 ； 困難腸梭菌

並列摘要

Many studies indicated that Clostridum difficile (C. difficile) can adhere on Caco-2 cell and damage the cell by secreting toxins. High C. cifficile toxin concentration may cause the cell death. Lactobacillus acidophilus GG (L. GG) and C. difficile were co-cultivated in the MRS medium in this study, the growth of C. difficile was inhibited by L. GG. After incubating for 16 hours, the growth rate of C. difficile was decreased and the CFU of C. difficile was decreased since 36 hours. When Caco-2 cells were infected by C. difficile, the lower TEER value revealed that the compact of Caco-2 cell membrane was ruined. However, the TEER value of the co-cultured sample was higher than that infection of C. difficile. The result indicated that C. difficile may damegae the monolayer membrane of Caco-2 cell and the probiotic L. GG can prevent the damage caused by C. difficile. This means that adsorption of L. GG may protect the intestines from antibio-associated pseudomembranous colitis caused by C. difficile. Growth of L. GG around C. difficile on the cell membrane surface or tight junction was observed.

並列關鍵字

Caco-2 cell ； Clostridium dfficile ； L. acidophilus GG

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