Development of the method for evaluating protective effect of food factors on THP-1-induced damage to human intestinal Caco-2 monolayers.
Immune cells located in the intestinal epithelium interact with intestinal epithelial cells via soluble factors. In this study, a new in vitro model using a coculture system was constructed to analyze the interaction between intestinal epithelial cells and macrophage-like cells. Human intestinal epithelial Caco-2 cells were differentiated on semipermeable membranes. Human monocytic THP-1 cells were differentiated to macrophage-like cells and then cocultured on the basolateral side of the Caco-2 cell monolayers. By coculturing for 48 hours, an increased release of lactate dehydrogenase from the Caco-2 cells and a decrease in the transepithelial electrical resistance of the monolayers were observed, suggesting that the coculture with THP-1 induced some disruption of the Caco-2 cells. This disruption was significantly suppressed by adding the anti-TNF-alpha antibody to the medium, suggesting that TNF-alpha secreted from THP-1 caused damage to the Caco-2 cells. It is also suggested that this phenomenon is similar to that observed with inflammatory bowel disease (IBD). The effects of food factors on the cells in this coculture system were examined. The disruption of the Caco-2 cell monolayers was significantly reduced by adding caffeine to the medium on the apical side. It is hoped that this coculture system will be a good model for the treatment of IBD.
Watanabe
Fumiko
F
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Satsu
Hideo
H
Mochizuki
Tetsunosuke
T
Nakano
Tomoko
T
Shimizu
Makoto
M
D002110:Caffeine / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000473:pathology*; D006801:Humans; D007413:Intestinal Mucosa / Q000473:pathology*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D047311:Luteolin / Q000494:pharmacology*; D008264:Macrophages / Q000502:physiology*; D009000:Monocytes; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism* / Q000494:pharmacology*
D014409:Tumor Necrosis Factor-alpha; D002110:Caffeine; D007770:L-Lactate Dehydrogenase; D047311:Luteolin