Release of Matrix Metalloproteinases-2 and 9 by S-Nitrosylated Caveolin-1 Contributes to Degradation of Extracellular Matrix in tPA-Treated Hypoxic Endothelial Cells (original) (raw)
Fig 2
ERK activation mediates MMP2 and 9 upregulation and their release in tPA and OGD-treated endothelial cells.
A) Gelatin zymography analysis showed that tPA alone significantly increased intracellular MMP2 and 9 levels, whereas their levels did not increase if tPA was administrated after OGD treatment (O+t). Upper panel: representative zymograms; Bottom panel: band intensity was quantitated and the data were expressed as mean ± SD, *p < 0.05, ANOVA, N = 3. Std: human MMP2 and 9 standards. B) Real time RT-PCR analysis showed that single tPA or tPA+OGD treatment (O+t) stimulated MMP2 and 9 mRNA expression in bEND3 cells. *p < 0.05, ANOVA, N = 4. C) tPA and OGD+tPA (O+t) treatment induced ERK phosphorylation (p-ERK). *p < 0.05, ANOVA, N = 5. D) U0126 completely blocked ERK activation. N = 3. E) TPA-induced MMP2 and 9 mRNA upregulation was inhibited by U0126. N = 3. F) The difference of MMP2 and 9 in cells among control (Ctrl), OGD, tPA, and OGD+tPA (O+t) was exterminated by U0126. Upper panel: representative zymograms; Bottom panel: band intensity was quantitated and the data were expressed as mean ± SD, ANOVA, N = 3. Std: human MMP2 and 9 standards. G) U0126 stopped OGD+tPA (O+t)-induced release of MMP2 and 9 in the media. Upper panel: representative zymograms; Bottom panel: band intensity was quantitated and the data were expressed as mean ± SD, ANOVA, N = 3. Std: human MMP2 and 9 standards.