Proteinase-activated receptor-2 mediates hyperresponsiveness in isolated guinea pig bronchi

The mast cell serine protease tryptase has been implicated as a critical mediator of airway hyperresponsiveness ill vitro and ill vivo. We have previously demonstrated that tryptase promotes hyperresponsiveness in isolated guinea pig bronchi. In this study, we have investigated the potential role of tryptase-mediated activation of proteinase-activated receptor-2 (PAR-2) in promoting airway hyperresponsiveness. Ex vivo exposure of guinea pig bronchi to the PAR-2 agonists H2N-Ser-Leu-Ile-Gly-Arg-Leu-CONH2 (SLIGRL) and t-cinnamoyl-H2N-Leu-Ile-Gly-Arg-Leu-O-CONH2 (t-c-LIGRLO) (0.1-10 muM) induced a concentration-dependent increase of contractile response to histamine. Treatment with 10 muM SLIGRL or t-c LIGRLO for 45 min increased subsequent responsiveness to histamine (0.3 mM) by 54 +/- 3% and 69 +/- 5%, respectively (P < 0.05 vs. control). In contrast, the PAR-1 agonist peptide H,N-Ser-Phe-Leu-Leu-Arg-Asn-CONH2 (SFLLRN) did not promote significant changes in the airway. Effects of the peptides were observed following at least a 30-min preincubation with the tissue. Coincubation with indomethacin or removal of epithelial cells is required for PAR-2-mediated hyperreactivity. The inactive analogue H2N-Leu-Ser-Ile-Gly-Arg-Leu-CONH2 (LISGRL; 10 muM) failed to promote hyperresponsiveness. Neuropeptide antagonists blocked the effect of the PAR-2 agonists. Selective antagonists of NK1 (L-703,606), NK2 (L-659,877). and CGRP (alphaCGRP 8-37) provided additive inhibition of PAR-2-mediated hyperreactivity. Pretreatment of bronchi with capsaicin (0.8 muM) also prevented the effects of SLIGRL. These results demonstrate the potential involvement of tryptase-mediated activation of PAR-2 in promoting airway hyperresponsiveness. These results further demonstrate that the PAR-2-mediated response involves a neurogenic mechanism involving neuropeptide release. (C) 2003 Elsevier Science Inc. All rights reserved.