Metformin influences cardiomyocyte cell death by pathways that are dependent and independent of caspase-3

• DOI: 10.1007/s00125-006-0338-9
• Publication Date: 2006-07-25T08:51:28Z
• Authors: D. An; G. Kewalramani; J. K. Y. Chan; D. Qi; S. Ghosh; T. Pulinilkunnil; A. Abrahani; S. M. Innis; B. Rodrigues
• Keywords: Male; 0301 basic medicine; Dose-Response Relationship, Drug; L-Lactate Dehydrogenase; Caspase 3; Blotting, Western; Fatty Acids; Palmitic Acid; Apoptosis; Hydrogen-Ion Concentration; Protein Serine-Threonine Kinases; Ceramides; Models, Biological; Metformin; 03 medical and health sciences; Animals; Myocytes, Cardiac; Lactic Acid; Phosphorylation; Glycolysis; Cells, Cultured; Acetyl-CoA Carboxylase

Metformin has been shown to increase fatty acid oxidation, an effect mediated by AMP activated protein kinase (AMPK). We hypothesised that metformin could prevent both caspase-3 activation and apoptosis when induced by palmitic acid.Cardiomyocytes were incubated with 1 mmol/l palmitic acid, in the absence or presence of metformin (1-5 mmol/l). Following 1 to 16 h, cell damage was evaluated by measuring lactate dehydrogenase released into the incubation medium, and Hoechst staining. To investigate the mechanism of metformin's effect on cardiomyocytes, substrate utilisation and phosphorylation of AMPK and acetyl-CoA carboxylase were measured. Intracellular mediators of apoptosis were also evaluated.Incubation of myocytes with palmitic acid for 16 h increased apoptosis, an effect that was partly blunted by 1 and 2 mmol/l metformin. This beneficial effect of metformin was associated with increased AMPK phosphorylation, palmitic acid oxidation and suppression of high-fat-induced increases in (1) long chain base biosynthesis protein 1 levels, (2) ceramide levels, and (3) caspase-3 activity. Unexpectedly, 5 mmol/l metformin dramatically increased apoptosis in myocytes incubated with high fat. This effect was associated with a robust increase in glycolysis, lactate accumulation, and a significant drop of pH in the myocyte incubation medium.Our study demonstrates that metformin reduces high-fat-induced cardiac cell death, probably through inhibition of ceramide synthesis. However, at high concentrations, metformin causes proton and lactate accumulation, leading to cell damage that is independent of caspase-3.