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

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 quest...

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Autores principales:	Elisa D’Alessandro, Billy Scaf, Chantal Munts, Arne van Hunnik, Christopher J. Trevelyan, Sander Verheule, Henri M. H. Spronk, Neil A. Turner, Hugo ten Cate, Ulrich Schotten, Frans A. van Nieuwenhoven
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	coagulation FXa cardiac fibroblasts inflammation PARs Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/ae3aa8565ee741448308919a364b907d
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spelling	oai:doaj.org-article:ae3aa8565ee741448308919a364b907d2021-11-25T17:09:49ZCoagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-110.3390/cells101129582073-4409https://doaj.org/article/ae3aa8565ee741448308919a364b907d2021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/2958https://doaj.org/toc/2073-4409Coagulation 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 <i>F2R</i> (PAR-1) and <i>F2RL1</i> (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 <i>Ccl2</i> (MCP-1; >30-fold at 4 h in atrial and ventricular CF) and <i>Il6</i> (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 <i>CCL2</i> (>3-fold) and <i>IL6</i> (>4-fold) at 4 h. Silencing of <i>F2R</i> (PAR-1 gene), but not <i>F2RL1</i> (PAR-2 gene), downregulated this effect. Selective activation of PAR-1 by TRAP-14 increased <i>CCL2</i> and <i>IL6</i> gene expression; this was prevented by <i>F2R</i> (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.Elisa D’AlessandroBilly ScafChantal MuntsArne van HunnikChristopher J. TrevelyanSander VerheuleHenri M. H. SpronkNeil A. TurnerHugo ten CateUlrich SchottenFrans A. van NieuwenhovenMDPI AGarticlecoagulation FXacardiac fibroblastsinflammationPARsBiology (General)QH301-705.5ENCells, Vol 10, Iss 2958, p 2958 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	coagulation FXa cardiac fibroblasts inflammation PARs Biology (General) QH301-705.5
spellingShingle	coagulation FXa cardiac fibroblasts inflammation PARs Biology (General) QH301-705.5 Elisa D’Alessandro Billy Scaf Chantal Munts Arne van Hunnik Christopher J. Trevelyan Sander Verheule Henri M. H. Spronk Neil A. Turner Hugo ten Cate Ulrich Schotten Frans A. van Nieuwenhoven Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
description	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 <i>F2R</i> (PAR-1) and <i>F2RL1</i> (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 <i>Ccl2</i> (MCP-1; >30-fold at 4 h in atrial and ventricular CF) and <i>Il6</i> (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 <i>CCL2</i> (>3-fold) and <i>IL6</i> (>4-fold) at 4 h. Silencing of <i>F2R</i> (PAR-1 gene), but not <i>F2RL1</i> (PAR-2 gene), downregulated this effect. Selective activation of PAR-1 by TRAP-14 increased <i>CCL2</i> and <i>IL6</i> gene expression; this was prevented by <i>F2R</i> (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.
format	article
author	Elisa D’Alessandro Billy Scaf Chantal Munts Arne van Hunnik Christopher J. Trevelyan Sander Verheule Henri M. H. Spronk Neil A. Turner Hugo ten Cate Ulrich Schotten Frans A. van Nieuwenhoven
author_facet	Elisa D’Alessandro Billy Scaf Chantal Munts Arne van Hunnik Christopher J. Trevelyan Sander Verheule Henri M. H. Spronk Neil A. Turner Hugo ten Cate Ulrich Schotten Frans A. van Nieuwenhoven
author_sort	Elisa D’Alessandro
title	Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
title_short	Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
title_full	Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
title_fullStr	Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
title_full_unstemmed	Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
title_sort	coagulation factor xa induces proinflammatory responses in cardiac fibroblasts via activation of protease-activated receptor-1
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ae3aa8565ee741448308919a364b907d
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Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

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