Figure 5

Effect of 3ABA on MMPs gelatinolytic activity. (a) Representative zymogram, showing MMP-2 and MMP-9 activity in medium from untreated cells (Control), in the presence of FGF2 (10 ng/ml), of FGF2 plus 3ABA (50 μM), and of 3ABA. HT1080 human fibrosarcoma cell conditioned medium was used as a marker of molecular weight. The areas of protease activity appeared as dark bands. (b) On the basis of image analysis of zymogram reported in Figure 5a, the percent amount of MMP-2 active form (MMP-2) over the summation (Σ) of pro-MMP-2 plus MMP-2 amounts was calculated. (c) Quantification of MMP-2 activity. Densitometric analysis of MMP-2 activity in medium from cells incubated in the absence (Control), in the presence of FGF2, of FGF2 plus 3ABA, and of 3ABA alone. Exposure of FGF2-stimulated HUVEC to 3ABA induced an increase in the active form of MMP-2 compared to both untreated cells (Control), and FGF2 stimulated cells. Values are expressed as percent of control and are mean ± SEM of three experiments. Multiple comparisons were performed by the Student-Newman-Keuls test, after Kruskal-Wallis analysis. (* p < 0.05 compared to control; # p < 0.05 compared to FGF2 treated cells).