Interleukin-1beta regulates cell proliferation and activity of extracellular matrix remodelling enzymes in cultured primary pig heart cells

Abstract

After myocardial infarction, elevated levels of interleukins (ILs) are found within the myocardial tissue and IL-1beta is considered to play a major role in tissue remodelling events throughout the body. In the study presented, we have established a cell culture model of primary pig heart cells to evaluate the effects of different concentrations of IL-1beta on cell proliferation as well as expression and activity of enzymes typically involved in tissue remodelling. Primary pig heart cell cultures were derived from three different animals and stimulated with recombinant pig IL-1beta. RNA expression was detected by RT-PCR, protein levels were evaluated by Western blotting, activity of matrix metalloproteinases (MMPs) was quantified by gelatine zymography and cell proliferation was measured using colorimetric MTS assays. Pig heart cells express receptors for IL-1 and application of IL-1beta resulted in a dose-dependent increase of cell proliferation (P<0.05 vs. control; 100ng/ml; 24h). Gene expression of caspase-3 was increased by IL-1beta (P<0.05 vs. control; 100ng/ml; 3h), and pro-caspase-3 but not active caspase was detected in lysates of pig heart cells by Western blotting. MMP-2 gene expression as well as enzymatic activities of MMP-2 and MMP-9 were increased by IL-1beta (P<0.05 vs. control; 100ng/ml; 3h for gene expression, 48 and 72h for enzymatic activities of MMP-2 and MMP-9, respectively). Our in vitro data suggest that IL-1beta plays a major role in the events of tissue remodelling in the heart. Combined with our recently published in vivo data (Meybohm et al., PLoS One, 2009), the results presented here strongly suggest IL-1beta as a key molecule guiding tissue remodelling events after myocardial infarction.