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).


METHODS
Evaluation of the effects of thrombin, thrombin receptor activator peptide 6 (TRAP-6), and D-phenylalanyl-1-propyl-L arginine chloromethyl ketone (PPACK) on the production of VEGF and MMPs by ESC.


METHODS
Research laboratory at the Oita University Medical School.


METHODS
Eight endometrial specimens in the secretory phase.


METHODS
ESC were incubated for 24 hours with thrombin, TRAP-6, and PPACK.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Furukawa
Yuichi
Y
Department of Obstetrics and Gynecology, Faculty of Medicine, Oita University, Oita, Japan.
Kawano
Yasushi
Y
Fukuda
Junichiro
J
Matsumoto
Harunobu
H
Narahara
Hisashi
H
D000328:Adult; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D015153:Blotting, Western; D002070:Butadienes / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004717:Endometrium / Q000187:drug effects / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D044463:Receptor, PAR-1 / Q000819:agonists / Q000378:metabolism*; D015398:Signal Transduction; D017154:Stromal Cells / Q000187:drug effects / Q000201:enzymology*; D013917:Thrombin / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D000590:Amino Acid Chloromethyl Ketones; D002070:Butadienes; D009570:Nitriles; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D044463:Receptor, PAR-1; C113580:U 0126; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C082835:thrombin receptor peptide (42-47); D020928:Mitogen-Activated Protein Kinases; D013917:Thrombin; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1; C040308:phenylalanyl-prolyl-arginine-chloromethyl ketone