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Serine proteases and protease-activated receptor 2 mediate the proinflammatory and algesic actions of diverse stimulants

Cattaruzza, F ; Amadesi, S ; Carlsson, J F ; Murphy, J E ; Lyo, V ; Kirkwood, K ; Cottrell, G S ; Bogyo, M ; Knecht, Wolfgang LU and Bunnett, N W (2014) In British Journal of Pharmacology 171(16). p.3814-3826
Abstract

BACKGROUND AND PURPOSE: Although serine proteases and agonists of protease-activated receptor 2 (PAR2) cause inflammation and pain, the spectrum of proteases that are activated by proinflammatory and algesic stimuli and their contribution to inflammatory pain are uncertain.

EXPERIMENTAL APPROACH: Enzymic assays and selective inhibitors were used to characterize protease activity in mice after intraplantar injections of formalin, bradykinin, PAR2 activating peptide (AP) or vehicle. The capacity of these proteases and of recombinant mouse trypsin 4 to cleave fragments of PAR2 and to activate PAR2 in cell lines was determined. Protease inhibitors and par2 (-/-) mice were used to assess the contributions of proteases and PAR2 to pain... (More)

BACKGROUND AND PURPOSE: Although serine proteases and agonists of protease-activated receptor 2 (PAR2) cause inflammation and pain, the spectrum of proteases that are activated by proinflammatory and algesic stimuli and their contribution to inflammatory pain are uncertain.

EXPERIMENTAL APPROACH: Enzymic assays and selective inhibitors were used to characterize protease activity in mice after intraplantar injections of formalin, bradykinin, PAR2 activating peptide (AP) or vehicle. The capacity of these proteases and of recombinant mouse trypsin 4 to cleave fragments of PAR2 and to activate PAR2 in cell lines was determined. Protease inhibitors and par2 (-/-) mice were used to assess the contributions of proteases and PAR2 to pain and inflammation.

KEY RESULTS: Intraplantar injection of formalin, bradykinin or PAR2-AP led to the activation of proteases that were susceptible to the serine protease inhibitor melagatran but resistant to soybean trypsin inhibitor (SBTI). Melagatran inhibited mouse trypsin 4, which degraded SBTI. Proteases generated in inflamed tissues cleaved PAR2-derived peptides. These proteases and trypsin 4 increased [Ca(2+) ]i in PAR2-transfected but not in untransfected cells, and melagatran suppressed this activity. Melagatran or PAR2 deletion suppressed oedema and mechanical hypersensitivity induced by intraplantar formalin, bradykinin and PAR2-AP, but had no effect on capsaicin-induced pain.

CONCLUSIONS AND IMPLICATIONS: Diverse proinflammatory and algesic agents activate melagatran-sensitive serine proteases that cause inflammation and pain by a PAR2-mediated mechanism. By inducing self-activating proteases, PAR2 amplifies and sustains inflammation and pain. Serine protease inhibitors can attenuate the inflammatory and algesic effects of diverse stimuli, representing a useful therapeutic strategy.

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author
; ; ; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Azetidines/pharmacology, Benzylamines/pharmacology, Bradykinin, Cell Line, Female, Foot, Formaldehyde, Inflammation/chemically induced, Male, Mice, Inbred C57BL, Mice, Transgenic, Oligopeptides, Pain/chemically induced, Receptor, PAR-2/agonists, Serine Proteases/metabolism, Serine Proteinase Inhibitors/pharmacology, Trypsin/metabolism
in
British Journal of Pharmacology
volume
171
issue
16
pages
3814 - 3826
publisher
Wiley
external identifiers
  • scopus:84903849154
  • pmid:24749982
ISSN
1476-5381
DOI
10.1111/bph.12738
language
English
LU publication?
no
id
4767c8c1-83d1-4ee1-a0a4-a94e21d4854a
date added to LUP
2020-07-22 14:06:01
date last changed
2024-03-20 14:23:40
@article{4767c8c1-83d1-4ee1-a0a4-a94e21d4854a,
  abstract     = {{<p>BACKGROUND AND PURPOSE: Although serine proteases and agonists of protease-activated receptor 2 (PAR2) cause inflammation and pain, the spectrum of proteases that are activated by proinflammatory and algesic stimuli and their contribution to inflammatory pain are uncertain.</p><p>EXPERIMENTAL APPROACH: Enzymic assays and selective inhibitors were used to characterize protease activity in mice after intraplantar injections of formalin, bradykinin, PAR2 activating peptide (AP) or vehicle. The capacity of these proteases and of recombinant mouse trypsin 4 to cleave fragments of PAR2 and to activate PAR2 in cell lines was determined. Protease inhibitors and par2 (-/-) mice were used to assess the contributions of proteases and PAR2 to pain and inflammation.</p><p>KEY RESULTS: Intraplantar injection of formalin, bradykinin or PAR2-AP led to the activation of proteases that were susceptible to the serine protease inhibitor melagatran but resistant to soybean trypsin inhibitor (SBTI). Melagatran inhibited mouse trypsin 4, which degraded SBTI. Proteases generated in inflamed tissues cleaved PAR2-derived peptides. These proteases and trypsin 4 increased [Ca(2+) ]i in PAR2-transfected but not in untransfected cells, and melagatran suppressed this activity. Melagatran or PAR2 deletion suppressed oedema and mechanical hypersensitivity induced by intraplantar formalin, bradykinin and PAR2-AP, but had no effect on capsaicin-induced pain.</p><p>CONCLUSIONS AND IMPLICATIONS: Diverse proinflammatory and algesic agents activate melagatran-sensitive serine proteases that cause inflammation and pain by a PAR2-mediated mechanism. By inducing self-activating proteases, PAR2 amplifies and sustains inflammation and pain. Serine protease inhibitors can attenuate the inflammatory and algesic effects of diverse stimuli, representing a useful therapeutic strategy.</p>}},
  author       = {{Cattaruzza, F and Amadesi, S and Carlsson, J F and Murphy, J E and Lyo, V and Kirkwood, K and Cottrell, G S and Bogyo, M and Knecht, Wolfgang and Bunnett, N W}},
  issn         = {{1476-5381}},
  keywords     = {{Animals; Azetidines/pharmacology; Benzylamines/pharmacology; Bradykinin; Cell Line; Female; Foot; Formaldehyde; Inflammation/chemically induced; Male; Mice, Inbred C57BL; Mice, Transgenic; Oligopeptides; Pain/chemically induced; Receptor, PAR-2/agonists; Serine Proteases/metabolism; Serine Proteinase Inhibitors/pharmacology; Trypsin/metabolism}},
  language     = {{eng}},
  number       = {{16}},
  pages        = {{3814--3826}},
  publisher    = {{Wiley}},
  series       = {{British Journal of Pharmacology}},
  title        = {{Serine proteases and protease-activated receptor 2 mediate the proinflammatory and algesic actions of diverse stimulants}},
  url          = {{http://dx.doi.org/10.1111/bph.12738}},
  doi          = {{10.1111/bph.12738}},
  volume       = {{171}},
  year         = {{2014}},
}