Figure 3

Effect of nintedanib on enzymatic activity (panel A-D) and secretion of matrix metalloproteinase (MMP)-2 (panel E, F) and tissue inhibitor of metalloproteinase (TIMP)-2 (panel G, H) by primary human lung IPF fibroblasts (A, C, E, G) and by non-fibrotic control cells (B, D, F, H). Fibroblasts were treated with increasing concentrations of nintedanib (0.001, 0.01, 0.1, and 1 μM) for 24 hours and conditioned cell medium was collected. (A, B) Representative gelatine-based zymograms of the effect of nintedanib on pro-MMP-2 enzymatic activity by IPF fibroblasts (A) and by control cells (B). Arrows indicate the migration position of purified pro-MMP-2. (C, D) Densitometric analysis of pro-MMP-2 enzymatic activity in primary human lung fibroblasts obtained from IPF lungs (C), and from non-fibrotic controls (D). Values are presented as mean ± SEM of independent experiments performed in 4 different cell lines, expressed as percentage of control (serum-free RPMI medium). Each experiment was performed at least in duplicates. *p < 0.05. The effect of nintedanib on the secretion of MMP-2 and TIMP-2 by IPF fibroblasts (E, G) and control cells (F, H) was assessed by ELISA. Values are presented as mean ± SEM of independent experiments performed in 4 different cell lines, expressed as percentage of control (serum-free RPMI medium). Each experiment was performed at least in duplicates. *p < 0.05.