PAR2-induced inflammatory responses in human kidney tubular epithelial cells

Vesey, David A., Suen, Jacky Y., Seow, Vernon, Lohman, Rink-Jan, Liu, Ligong, Gobe, Glenda C., Johnson, David W. and Fairlie, David P. (2013) PAR2-induced inflammatory responses in human kidney tubular epithelial cells. American Journal of Physiology - Renal Physiology, 304 6: F737-F750. doi:10.1152/ajprenal.00540.2012

Author Vesey, David A.
Suen, Jacky Y.
Seow, Vernon
Lohman, Rink-Jan
Liu, Ligong
Gobe, Glenda C.
Johnson, David W.
Fairlie, David P.
Title PAR2-induced inflammatory responses in human kidney tubular epithelial cells
Journal name American Journal of Physiology - Renal Physiology   Check publisher's open access policy
ISSN 1931-857X
Publication date 2013-03-15
Year available 2013
Sub-type Article (original research)
DOI 10.1152/ajprenal.00540.2012
Volume 304
Issue 6
Start page F737
End page F750
Total pages 14
Place of publication Bethesda, MD, United States
Publisher American Physiological Society
Collection year 2014
Language eng
Formatted abstract
Protease-activated receptor-2 (PAR2) is a G protein-coupled receptor abundantly expressed in the kidney. The aim of this study was to profile inflammatory gene and protein expression induced by PAR2 activation in human kidney tubular epithelial cells (HTEC). A novel PAR2 antagonist, GB88, was used to confirm agonist specificity. Intracellular Ca2+ (iCa2+) mobilization, confocal microscopy, gene expression profiling, qRTPCR, and protein expression were used to characterize PAR2 activation. PAR2 induced a pronounced increase in iCa2+ concentration that was blocked by the PAR2 antagonist. Treatment with SLIGKV-NH2 at the apical or basolateral cell surface for 5 h induced expression of a range of inflammatory genes by greater than fourfold, including IL-1β, TRAF1, IL-6, and MMP-1, as assessed by cDNA microarray and qRTPCR analysis. Using antibody arrays, GM-CSF, ICAM-1, TNF-α, MMP-1, and MMP-10 were among the induced proteins secreted. Cytokine-specific ELISAs identified three- to sixfold increases in GM-CSF, IL-6, IL-8, and TNF-α, which were blocked by GB88 and protein kinase C inhibitors. Treatment of cells at the basolateral surface induced more potent inflammatory responses, with release of MCP-1 and fibronectin to the apical and basolateral compartments; apical treatment only increased secretion of these factors to the apical compartment. PAR2 activation at the basolateral surface dramatically reduced transepithelial electrical resistance (TEER) whereas apical treatment had no effect. There was very little leakage (<5%) of peptides across the cell monolayer (liquid chromatography-mass spectrometry). In summary, SLIGKV-NH2 induced robust proinflammatory responses in HTEC that were antagonized by GB88. These results suggest that PAR2 antagonists could be useful disease-modifying, anti-inflammatory agents in kidney disease.
Keyword Protease-activated receptor-2
Kidney tubule
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
School of Medicine Publications
Institute for Molecular Bioscience - Publications
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