Intestinal epithelial cells can be induced to secrete the chemokine interleukin (IL)-8 during inflammation. The PAR-2 receptor is believed to play a proinflammatory role and is expressed in gut epithelial cells. The aim was to investigate PAR-2 signaling in Caco-2 intestinal epithelial cells, with respect to chemokine secretion. Activation of PAR-2 by high concentrations of the synthetic activating peptide (SLIGKV) did not induce secretion of IL-8, in contrast to stimulation with IL-1beta. However, upon simultaneous treatment with activating peptide and IL-1beta, a potentiating effect of PAR-2 stimulation was seen, resulting in a fivefold increase of IL-8. Available data suggest that NF-kappaB activation is required for IL-8 gene expression. Unlike IL-1beta, PAR-2 stimulation did not activate NF-kappaB, which may explain the lack of IL-8 expression. However, PAR-2 stimulation led to rapid phosphorylation of two MAP kinases, p38 MAPK and ERK1/2. ERK1/2 is known to activate the transcription factor AP-1, also involved in upregulation of IL-8 gene transcription. Inhibition of p38 MAPK led to decreased IL-8 following stimulation with IL-1beta and/or activating peptide. These results suggest that maximal IL-8 expression requires coordination of several signaling pathways. Thus, identifying antagonists to the PAR-2 receptor may be beneficial by inhibiting potentiation of a proinflammatory response, through inhibition of p38 and ERK MAP kinases.