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Am J Physiol Cell Physiol 278: C463-C472, 2000;
0363-6143/00 $5.00
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Vol. 278, Issue 3, C463-C472, March 2000

TNF-alpha potentiates the ion secretion induced by muscarinic receptor activation in HT29cl.19A cells

Judith C. J. Oprins, Helen P. Meijer, and Jack A. Groot

Institute for Neurobiology, Biological Faculty, University of Amsterdam, 1098 SM Amsterdam, The Netherlands

Chronic gastrointestinal diseases such as ulcerative colitis and Crohn's disease are characterized by severe diarrhea. Mucosal biopsies of these patients show enhanced levels of cytokines, secreted by infiltrated inflammatory cells. In this study, we investigated the effect of the cytokine tumor necrosis factor-alpha (TNF-alpha ) on ion secretion in human intestinal epithelial cells. The conventional microelectrode technique in the cell line HT29cl.19A was used, which allows for simultaneous measurements of transepithelial potential difference and intracellular potential difference across the apical membrane. Preincubation (2-78 h) with 10 ng/ml TNF-alpha did not change basal secretory activity. However, the secretory response to the muscarinic receptor agonist carbachol was strongly increased after exposure to TNF-alpha . Application of the protein kinase C (PKC) inhibitor GF 109203X (bisindolylmaleimide I) inhibited the response to carbachol as well as the TNF-alpha -potentiated response, indicating that PKC mediates the effect of carbachol in this cell line. Propranolol, a substance that inhibits the phospholipase D (PLD) pathway, strongly reduced the response to muscarinic stimulation and its potentiation by TNF-alpha . The results indicate that activation of PLD is involved in ion secretion induced by muscarinic receptor activation and that TNF-alpha can potentiate this pathway.

phospholipase D; protein kinase C; intestinal epithelia; carbachol; cytokine; tumor necrosis factor-alpha


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