Thrombospondin-1 regulation of smooth muscle cell chemotaxis is extracellular signal-regulated protein kinases 1/2 dependent
| Autor: | V, Gahtan, X J, Wang, A I, Willis, G P, Tuszynski, B E, Sumpio |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3 Dose-Response Relationship Drug Chemotaxis Protein-Tyrosine Kinases Muscle Smooth Vascular Thrombospondin 1 Focal Adhesion Protein-Tyrosine Kinases Calcium-Calmodulin-Dependent Protein Kinases Animals Cattle Mitogen-Activated Protein Kinases Cell Adhesion Molecules Cells Cultured |
| Zdroj: | Surgery. 126(2) |
| ISSN: | 0039-6060 |
| Popis: | Thrombospondin-1 (TSP-1), an extracellular matrix protein, induces vascular smooth muscle cell (VSMC) chemotaxis. We hypothesized that extracellular signal-regulated protein kinases 1/2 (ERK1/2), a pathway of the mitogen activated protein kinase (MAPK) family, is important in TSP-1-induced VSMC chemotaxis.A modified Boyden chamber was used to assess chemotaxis. First, a concentration curve was performed to determine the level for optimal TSP-1-induced chemotaxis. Then quiescent VSMCs were preincubated (30 minutes) in serum-free medium, dimethyl sulfoxide (the inhibitor vehicle), or PD98059 (10 mumol/L, an upstream inhibitor of ERK1/2). VSMCs (50,000 cells/well) with the appropriate preincubation were placed in the top chamber. The bottom chamber contained TSP-1 (20 micrograms/mL) or serum-free medium. Results were recorded as cells/5 fields (400x). Then quiescent VSMCs were exposed to TSP-1 (20 micrograms/mL) for 0, 1, 5, 10, 30, 120, or 300 minutes. Platelet-derived growth factor (10 ng/mL) was the positive control for ERK1/2 activation. Western blot analysis was performed for activated ERK1/2. All comparisons were made by a paired t test (n = 3).TSP-1-induced chemotaxis peaks by a concentration of 20 micrograms/mL. PD98059 inhibited TSP-1-induced chemotaxis (P.05). ERK1/2 was activated by TSP-1-stimulated VSMCs.TSP-1-stimulated VSMCs activated ERK1/2. An ERK1/2 inhibitor abolished chemotaxis, suggesting the functional importance of MAPK in TSP-1-induced VSMC chemotaxis. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect