Journal Article

Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

ten Cate, Hugo, Trevelyan, Christopher J., van Hunnik, Arne, D’Alessandro, Elisa, Schotten, Ulrich, Scaf, Billy, Munts, Chantal, Verheule, Sander, Spronk, Henri M. H., van Nieuwenhoven, Frans A., Turner, Neil A.

2021

Overview

Coagulation factor (F) Xa induces proinflammatory responses through activation of protease-activated receptors (PARs). However, the effect of FXa on cardiac fibroblasts (CFs) and the contribution of PARs in FXa-induced cellular signalling in CF has not been fully characterised. To answer these questions, human and rat CFs were incubated with FXa (or TRAP-14, PAR-1 agonist). Gene expression of pro-fibrotic and proinflammatory markers was determined by qRT-PCR after 4 and 24 h. Gene silencing of F2R (PAR-1) and F2RL1 (PAR-2) was achieved using siRNA. MCP-1 protein levels were measured by ELISA of FXa-conditioned media at 24 h. Cell proliferation was assessed after 24 h of incubation with FXa ± SCH79797 (PAR-1 antagonist). In rat CFs, FXa induced upregulation of Ccl2 (MCP-1; >30-fold at 4 h in atrial and ventricular CF) and Il6 (IL-6; ±7-fold at 4 h in ventricular CF). Increased MCP-1 protein levels were detected in FXa-conditioned media at 24 h. In human CF, FXa upregulated the gene expression of CCL2 (>3-fold) and IL6 (>4-fold) at 4 h. Silencing of F2R (PAR-1 gene), but not F2RL1 (PAR-2 gene), downregulated this effect. Selective activation of PAR-1 by TRAP-14 increased CCL2 and IL6 gene expression; this was prevented by F2R (PAR-1 gene) knockdown. Moreover, SCH79797 decreased FXa-induced proliferation after 24 h. In conclusion, our study shows that FXa induces overexpression of proinflammatory genes in human CFs via PAR-1, which was found to be the most abundant PARs isoform in this cell type.

• Publisher: MDPI AG,MDPI
• Subject: Adult / Animals / Biopsy / cardiac fibroblasts / Cattle / Cell Proliferation / Chemokine CCL2 - genetics / Chemokine CCL2 - metabolism / Coagulation / Coagulation factor Xa / coagulation FXa / Factor Xa - metabolism / Fibroblasts / Fibroblasts - metabolism / Fibroblasts - pathology / Gene expression / Gene silencing / Growth factors / Heart / Heart Atria - pathology / Heart attacks / Humans / Inflammation / Inflammation - pathology / Interleukin 6 / Interleukin-6 - genetics / Interleukin-6 - metabolism / Monocyte chemoattractant protein 1 / Myocardium - metabolism / Par-1 protein / Par-2 protein / PARs / Protein folding / Proteinase-activated receptor 1 / Rats / Rats, Wistar / Receptor, PAR-1 - agonists / Receptor, PAR-1 - genetics / Receptor, PAR-1 - metabolism / siRNA / Thrombin - metabolism / Tumor necrosis factor-TNF / Up-Regulation - genetics / Ventricle