Pyruvate reverses fatty-acid-induced depression of ventricular function and calcium overload after hypothermia in guinea pig hearts

Objective: High levels of free fatty acids have been shown to impair mechanical recovery and calcium homeostasis of isolated rat hearts following hypothermic perfusion. The objective of the present study was to investigate whether inhibition of fatty acid oxidation through activation of pyruvate dehydrogenase by millimolar concentrations of pyruvate could influence functional recovery and Ca2+ homeostasis after a hypothermic insult. Methods: Ventricular function and myocardial calcium ([Ca](total)) were measured in 3 different groups of Langendorff-perfused guinea pig hearts exposed to 40 min hypothermic (15 degrees C) perfusion, followed by 30 min rewarming at 37 degrees C. The hearts were perfused with either 11.1 mM glucose (G), glucose and 1.2 mM palmitate (GPI, or glucose, palmitate and 5 mM pyruvate (GPP) as energy substrates. Results: All groups showed marked elevations in [Ca](total) during hypothermia (from 0.6-0.7 mu mol . g . dry wt(-1) to 9.3-12.2 mu mol . g dry wt(-1) at 40 min hypothermia, P < 0.05), associated with a pronounced increase in left ventricular end-diastolic pressure (LVEDP from 0-2 to 50-60 mmHg). Following rewarming, GP-perfused hearts showed significantly lower recovery of mechanical function compared to both G- and GPP-perfused hearts (% recovery of left ventricular developed pressure: 27 +/- 8 vs. 62 +/- 3 and 62 +/- 8%, respectively, P < 0.05). The reduced mechanical recovery of GP-perfused hearts was associated with elevated [Ca](total). In separate experiments we found that addition of 1.2 mM palmitate reduced glucose oxidation ([C-14]glucose) from 1.77 +/- 0.28 mu mol . min(-1). g dry wt(-1) (G-perfused hearts) to 0.15 +/- 0.04 mu mol . min(-1). g dry wt(-1) (GP-perfused hearts, P < 0.05), implying that fatty acids had become the major substrate for oxidative phosphorylation. Fatty acid oxidation was, however, less pronounced after further addition of 5 mM pyruvate. Thus, palmitate oxidation ([H-3]palmitate) was more than 40% lower in GPP-perfused than in GP-perfused hearts (0.83 +/- 0.22 vs. 1.41 +/- 0.12 mu mol . min(-1). g . dry wt(-1), P < 0.05). Conclusions: The present results demonstrate impaired ventricular function and calcium homeostasis after hypothermia in guinea pig hearts perfused with fatty acids in addition to glucose, as compared to hearts perfused with glucose alone. Futhermore, we show that these unfavourable effects of fatty acids can be overcome by an exogenous supply of pyruvate.