Levosimendan inhibits interleukin-1β-induced cell migration and MMP-9 secretion in rat cardiac fibroblasts

Abstract

Cell migration and matrix metalloproteinases (MMPs) secretion in cardiac fibroblasts are important processes in the cardiac remodeling during the development of cardiac diseases and are regulated by proinflammatory cytokines. Although levosimendan, a novel inotropic agent, is expected to have some beneficial influences on preventing cardiac remodeling, its effects on proinflammatory cytokines-induced functional changes in cardiac fibroblasts have not been clarified. Therefore, we investigated the effects of levosimendan on interleukin (IL)-1β−induced MMP-9 secretion and migration in adult rat cardiac fibroblasts. Primary cardiac fibroblasts were isolated from adult male Wistar rats. MMP-9 secretion in culture medium and extracellular signal-regulated kinase (ERK) phosphorylation in cell lysate were measured by using Western blotting. Gelatin zymography was performed to measure activity of secreted MMP-9. MMP-9 mRNA expression in the cell was measured by using reverse transcription polymerase chain reaction. Boyden chamber assay was performed for detection of migration. Levosimendan (3–100μM) concentration-dependently inhibited IL-1β (4ng/ml)-induced MMP-9 secretion, activity and mRNA expression. Levosimendan inhibited IL-1β (4ng/ml)-induced ERK phosphorylation. Levosimendan (10 and 100μM) inhibited IL-1β-induced migration, and CTTHWGFTLC peptide (10μM), an MMP inhibitor, or PD98059 (50μM), an ERK inhibitor, also suppressed it. The present study for the first time demonstrated in adult rat cardiac fibroblasts that levosimendan inhibits IL-1β-induced migration at least partly through the inhibition of MMP-9 secretion via suppressing ERK phosphorylation.