PAR-1 and PAR-2 Expression Is Enhanced in Inflamed Odontoblast Cells

PAR-1 and PAR-2 Expression Is Enhanced in Inflamed Odontoblast Cells

Author Alvarez, M. M. P. Autor UNIFESP Google Scholar
Moura, G. E. Autor UNIFESP Google Scholar
Machado, M. F. M. Google Scholar
Viana, G. M. Autor UNIFESP Google Scholar
de Souza Costa, C. A. Google Scholar
Tjaderhane, L. Google Scholar
Nader, H. B. Autor UNIFESP Google Scholar
Tersariol, I. L. S. Autor UNIFESP Google Scholar
Nascimento, F. D. Google Scholar
Abstract Protease-activated receptors (PARs) are G protein-coupled receptors, which are activated by proteolytical cleavage of the amino-terminus and act as sensors for extracellular proteases. We hypothesized that PAR-1 and PAR-2 can be modulated by inflammatory stimulus in human dental pulp cells. PAR-1 and PAR-2 gene expression in human pulp tissue and MDPC-23 cells were analyzed by quantitative polymerase chain reaction. Monoclonal PAR-1 and PAR-2 antibodies were used to investigate the cellular expression of these receptors using Western blot, flow cytometry, and confocal microscopy in MDPC-23 cells. Immunofluorescence assays of human intact and carious teeth were performed to assess the presence of PAR-1 and PAR-2 in the dentin-pulp complex. The results show for the first time that human odontoblasts and MDPC-23 cells constitutively express PAR-1 and PAR-2. PAR-2 activation increased significantly the messenger RNA expression of matrix metalloproteinase (MMP)-2, MMP-9, MMP-13, and MMP-14 in MDPC-23 cells (P < 0.05), while the expression of these enzymes decreased significantly in the PAR-1 agonist group (P < 0.05). The high-performance liquid chromatography and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis showed the presence of MMP-13 activity cleaving PAR-1 at specific, noncanonical site TLDPRS42 down arrow(FLL)-L-43 in human dental pulp tissues. Also, we detected a presence of a trypsin-like activity cleaving PAR-2 at canonical site SKGR(20)down arrow S(21)LIGRL in pulp tissues. Confocal microscopy analysis of human dentin-pulp complex showed intense positive staining of PAR-1 and PAR-2 in the odontoblast processes in dentinal tubules of carious teeth compared to intact ones. The present results support the hypothesis of activation of the upregulated PAR-1 and PAR-2 by endogenous proteases abundant during the inflammatory response in dentin-pulp complex.
Keywords proteinase-activated receptor (PAR)
dentin-pulp complex
proteolytic enzymes
matrix metalloproteinases
xmlui.dri2xhtml.METS-1.0.item-coverage Thousand Oaks
Language English
Sponsor FAPESP, Brazil
CAPES, Brazil
CNPq, Brazil
Grant number FAPESP: 13/05822-9
FAPESP: 15/03964-6
Date 2017
Published in Journal Of Dental Research. Thousand Oaks, v. 96, n. 13, p. 1518-1525, 2017.
ISSN 0022-0345 (Sherpa/Romeo, impact factor)
Publisher Sage Publications Inc
Extent 1518-1525
Access rights Closed access
Type Article
Web of Science ID WOS:000415925000008

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