The Effects of Protease-Activated Receptor 4 Stimulation on Vascular Smooth Muscle Cell Migration and Phosphorylation of ERK
Protease-Activated Receptor 4 is a G-protein coupled receptor that is present in Vascular Smooth Muscle Cells. It is activated by thrombolytic cleavage and involved in multiple cellular functions including inflammation, cell proliferation, cell aggregation, and platelet activation. It is also believed that PAR4 may play a role in the development of Cardiovascular Disease. PAR4 creates a great deal of research interest because there is still a great deal to learn about the receptor’s mechanisms of action. However, it is believed that there are possible therapeutic benefits in the future development of a safe PAR4 antagonist for patients being treated for cardiovascular disease. The purpose of this study was to determine the role PAR4 plays in cell migration and phosphorylation of ERK. Experiments were performed using rat aortic smooth muscle cells. Cells were placed on six-well plates and serum starved for 24 hours. A 24-well cell migration assay kit was used to determine cell migration when stimulated with PAR4 agonist (AY-NH2) for 15 minutes, 2 hours, and 24 hours. Cells were then examined using a plate reader at OD 560 nm. Results showed that cells stimulated with PAR4 agonist for 15 minutes and 2 hours had a higher percentage of migration than cells that were not stimulated with agonist. However, cells stimulated with agonist for 24 hours showed a lower migration than those not stimulated at all. In a second experiment, cells in a 6-well plate were stimulated with PAR4 agonist for 0, 1, 2, 5, 15, and 30 minutes. A Western Blot procedure was performed to determine the level of ERK phosphorylation. Results showed bands around 50 kDa. This signifies that PAR4 stimulation causes phosphorylation of ERK 1/2. Overall, the results of this study showed that PAR4 stimulation causes increased cell migration and caused phosphorylation of ERK 1/2.
"The Effects of Protease-Activated Receptor 4 Stimulation on Vascular Smooth Muscle Cell Migration and Phosphorylation of ERK"
ETD Collection for Tennessee State University.