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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MDPI AG
2021-10-01
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T05:37:38Z |
| format | Article |
| id | doaj.art-ae3aa8565ee741448308919a364b907d |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T05:37:38Z |
| publishDate | 2021-10-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-ae3aa8565ee741448308919a364b907d2023-11-22T22:49:15ZengMDPI AGCells2073-44092021-10-011011295810.3390/cells10112958Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1Elisa D’Alessandro0Billy Scaf1Chantal Munts2Arne van Hunnik3Christopher J. Trevelyan4Sander Verheule5Henri M. H. Spronk6Neil A. Turner7Hugo ten Cate8Ulrich Schotten9Frans A. van Nieuwenhoven10Departments of Biochemistry and Internal Medicine, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6220 MD Maastricht, The NetherlandsDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsDiscovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UKDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsDepartments of Biochemistry and Internal Medicine, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6220 MD Maastricht, The NetherlandsDiscovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UKDepartments of Biochemistry and Internal Medicine, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6220 MD Maastricht, The NetherlandsDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsDepartment of Physiology, Cardiovascular Research Institute Maastricht, Maastricht University Medical Center, 6200 MD Maastricht, The NetherlandsCoagulation 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.https://www.mdpi.com/2073-4409/10/11/2958coagulation FXacardiac fibroblastsinflammationPARs |
| spellingShingle | 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 Cells coagulation FXa cardiac fibroblasts inflammation PARs |
| title | 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_short | 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 |
| topic | coagulation FXa cardiac fibroblasts inflammation PARs |
| url | https://www.mdpi.com/2073-4409/10/11/2958 |
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