Abstract
Objective
This study aims to investigate the role of protease-activated receptor (PAR) 2 and mast cell (MC) tryptase in LPS-induced lung inflammation and neutrophil recruitment in the lungs of C57BL/6 mice.
Methods
C57BL/6 mice were pretreated with the PAR2 antagonist ENMD-1068, compound 48/80 or aprotinin prior to intranasal instillation of MC tryptase or LPS. Blood leukocytes, C-X-C motif chemokine ligand (CXCL) 1 production leukocytes recovered from bronchoalveolar lavage fluid (BALF), and histopathological analysis of the lung were evaluated 4 h later. Furthermore, we performed experiments to determine intracellular calcium signaling in RAW 264.7 cells stimulated with LPS in the presence or absence of a protease inhibitor cocktail or ENMD-1068 and evaluated PAR2 expression in the lungs of LPS-treated mice.
Results
Pharmacological blockade of PAR2 or inhibition of proteases reduced neutrophils recovered in BALF and LPS-induced calcium signaling. PAR2 blockade impaired LPS-induced lung inflammation, PAR2 expression in the lung and CXCL1 release in BALF, and increased circulating blood neutrophils. Intranasal instillation of MC tryptase increased the number of neutrophils recovered in BALF, and MC depletion with compound 48/80 impaired LPS-induced neutrophil migration.
Conclusion
Our study provides, for the first time, evidence of a pivotal role for MCs and MC tryptase in neutrophil migration, lung inflammation and macrophage activation triggered by LPS, by a mechanism dependent on PAR2 activation.
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Change history
07 August 2020
The original article can be found online.
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Acknowledgements
This work was supported by Fundação de Amparo à Pesquisa de Minas Gerais (FAPEMIG/Brazil), Grant Number PPM-00593-16. A.D.A. was a graduate student fellow from the Conselho Nacional de Pesquisa e Desenvolvimento (CNPq/Brazil), A.C.M.L.F. and A.B. are graduate students from CNPq. L.O.A., M.F.L. and G.D.C. are research fellows from the CNPq. I.S.S. and and R.M.F. are graduate student from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/Brazil)
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ADA: conceptualization, investigation, formal analysis, writing original draft; ISS, WFB, ACMLF, RMF, AB: investigation; LOA, MFL: resources; GDC: resources, formal analysis; AK: supervision, project administration, funding acquisition, writing review and editing.
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de Almeida, A.D., Silva, I.S., Fernandes-Braga, W. et al. A role for mast cells and mast cell tryptase in driving neutrophil recruitment in LPS-induced lung inflammation via protease-activated receptor 2 in mice. Inflamm. Res. 69, 1059–1070 (2020). https://doi.org/10.1007/s00011-020-01376-4
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DOI: https://doi.org/10.1007/s00011-020-01376-4