Intestinal inflammation increases the expression of the P2Y6 receptor on epithelial cells and the release of CXC chemokine ligand 8 by UDP.
Epithelial cells participate in the immune response of the intestinal mucosa. Extracellular nucleotides have been recognized as inflammatory molecules. We investigated the role of extracellular nucleotides and their associated P2Y receptors in the secretion of cytokines by epithelial cells. The effect of intestinal inflammation on P2Y(6) receptor expression was determined by PCR in the mouse, rat, and human. Localization of the P2Y(6) receptor was determined by immunofluorescence microscopy in the colon of normal and dextran sulfate sodium-treated mice. The effect of P2Y(6) activation by UDP on cytokine expression and release by epithelial cells was determined using a combination of Western blots, luciferase assays, RT-PCR, cytokine Ab arrays, and ELISA. Inflammation up-regulates P2Y(2) as well as P2Y(6) receptor expression in the mucosa of the colon of colitic mice. In vitro, we demonstrated that UDP could be released by Caco-2/15 cells. We have confirmed the increased expression of P2Y(6) by challenging intestinal epithelial cell-6 and Caco-2/15 cells with TNF-alpha and IFN-gamma and showing that stimulation of epithelial cells by UDP results in an increased expression and release of CXCL8 by an ERK1/2-dependent mechanism. The increase in CXCL8 expression was associated with a transcriptional activation by the P2Y(6) receptor. This study is the first report demonstrating the implication of P2Y receptors in the inflammatory response of intestinal epithelial cells. We show for the first time that P2Y(6), as well as P2Y(2), expression is increased by the stress associated with intestinal inflammation. These results demonstrate the emergence of extracellular nucleotide signaling in the orchestration of intestinal inflammation.
Grbic
Djordje M
DM
Canadian Institutes of Health Research Team on the Digestive Epithelium, Département d'Anatomie et de Biologie Cellulaire, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, 3001 12th Avenue North, Sherbrooke, Québec, Canada.
Degagné
Emilie
E
Langlois
Christine
C
Dupuis
Andrée-Anne
AA
Gendron
Fernand-Pierre
FP
D000818:Animals; D018938:Caco-2 Cells; D002460:Cell Line; D003092:Colitis / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D007249:Inflammation / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism*; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D018384:Oxidative Stress / Q000276:immunology; D010766:Phosphorylation; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D058473:Receptors, Purinergic P2Y2; D015854:Up-Regulation / Q000276:immunology*; D014530:Uridine Diphosphate / Q000378:metabolism / Q000502:physiology*; D014544:Uridine Triphosphate / Q000502:physiology
D018836:Inflammation Mediators; D016209:Interleukin-8; C546385:P2RY2 protein, human; C546386:P2ry2 protein, mouse; C546387:P2ry2 protein, rat; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; C099845:purinoceptor P2Y6; D014530:Uridine Diphosphate; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D014544:Uridine Triphosphate