The production of vascular endothelial growth factor and metalloproteinase via protease-activated receptor in human endometrial stromal cells

Objective: To measure the levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs) induced by thrombin in endometrial stromal cells (ESC). Design: Evaluation of the effects of thrombin, thrombin receptor activator peptide 6 (TRAP-6), and D-phenylalanyl-1-ptopyl-L arginine chloromethyl ketone (PPACK) on the production of VEGF and MMPs by ESC. Setting: Research laboratory at the Oita University Medical School. Patient(S): Eight endometrial specimens in the secretory phase. Intervention(S): ESC were incubated for 24 hours with thrombin, TRAP-6, and PPACK. Main Outcome Measure(s): The levels of VEGF, MMP-1, and active MMP-2 were measured by enzyme-linked immunosorbent assay (ELISA). The presence of protease-activated receptor-1 (PAR-1) and activation of mitogen-activated protein (MAP) kinase were detected by Western blot analysis. Result(s): Following stimulation by thrombin and TRP-6, the production of VEGF, MMP-1, and active MMP-2 statistically significantly increased; U0126 and PPACK statistically significantly suppressed the increases in the production of VEGF, MMP-1, and active MMP-2 induced by thrombin and TRAP-6. Activity by MAP kinase was induced by treatment with thrombin and TRAP-6 and was suppressed by PPACK. Conclusion(S): The results suggest that thrombin stimulates the production of VEGF and MMPs by a mechanism involving the MAP kinase system. The increases in VEGF and MMPs may contribute to neovascularization, which promotes the proliferation of endometrium and placentation. (Fertil Steril(R) 2009;91:535-41. (C)2009 by American Society for Reproductive Medicine.)