Expression of Matrix Metalloproteases by Fibrocytes
| Autor: | Victor Ruiz, Silvia Reyes, Carolina Garcia-de-Alba, Moisés Selman, Yolanda González, Annie Pardo, Carina Becerril, Jorge Garcia-Alvarez |
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| Rok vydání: | 2010 |
| Předmět: |
Pulmonary and Respiratory Medicine
Pathology medicine.medical_specialty Pulmonary Fibrosis Blotting Western Matrix metalloproteinase Critical Care and Intensive Care Medicine Polymerase Chain Reaction Immunophenotyping Flow cytometry Transforming Growth Factor beta1 Cell Movement Fibrocyte medicine Humans Progenitor cell Cells Cultured Microscopy Confocal medicine.diagnostic_test business.industry Mesenchymal stem cell Cell migration Fibroblasts Matrix Metalloproteinases Extracellular Matrix Cell biology Matrix Metalloproteinase 8 Matrix Metalloproteinase 7 Collagen Stem cell Cell Migration Assays business Homing (hematopoietic) |
| Zdroj: | American Journal of Respiratory and Critical Care Medicine. 182:1144-1152 |
| ISSN: | 1535-4970 1073-449X |
| DOI: | 10.1164/rccm.201001-0028oc |
| Popis: | Fibrocytes are progenitor cells characterized by the simultaneous expression of mesenchymal, monocyte, and hematopoietic stem cell markers. We previously documented their presence in lungs of patients with idiopathic pulmonary fibrosis. However, the mechanisms involved in their migration, subsequent homing, and local role remain unclear. Matrix metalloproteinases (MMPs) facilitate cell migration and have been implicated in the pathogenesis of pulmonary fibrosis.To evaluate the expression and role of matrix metalloproteinases in human fibrocytes.Fibrocytes were purified from CD14(+) monocytes and cultured for 8 days; purity of fibrocyte cultures was 95% or greater as determined by flow cytometry. Conditioned media and total RNA were collected and the expression of MMP-1, MMP-2, MMP-7, MMP-8, and MMP-9 was evaluated by real-time polymerase chain reaction. Protein synthesis was examined using a Multiplex assay, Western blot, fluorescent immunocytochemistry, and confocal microscopy. MMP-2 and MMP-9 enzymatic activities were evaluated by gelatin zymography. Migration was assessed using collagen I-coated Boyden chambers. Stromal cell-derived factor-1α and platelet-derived growth factor-B were used as chemoattractant with or without a specific MMP-8 inhibitor.Fibrocytes showed gene and protein expression of MMP-2, MMP-9, MMP-8, and MMP-7. MMP-2 and MMP-9 enzymatic activities were also demonstrated by gelatin zymography. Likewise, we found colocalization of MMP-8 and MMP-7 with type I collagen in fibrocytes. Fibrocyte migration toward platelet-derived growth factor-B or Stromal cell-derived factor-1α in collagen I-coated Boyden chambers was significantly reduced by a specific MMP-8 inhibitor.Our findings reveal that fibrocytes express a variety of MMPs and that MMP-8 actively participates in the process of fibrocyte migration. |
| Databáze: | OpenAIRE |
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