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Warm ischemic preconditioning improves mitochondrial redox balance during and after mild hypothermic ischemia in guinea pig isolated hearts

Anesthesiology Research Laboratory, Departments of ¹Anesthesiology and ²Physiology, Medical College of Wisconsin, ³Department of Biomedical Engineering, Marquette University, and ⁴Research Service, Veterans Affairs Medical Center, Milwaukee, Wisconsin

Submitted 5 November 2004 ; accepted in final form 8 January 2005

Ischemic preconditioning (IPC) induces distinctive changes in mitochondrial bioenergetics during warm (37°C) ischemia and improves function and tissue viability on reperfusion. We examined whether IPC before 2 h of hypothermic (27°C) ischemia affords additive cardioprotection and improves mitochondrial redox balance assessed by mitochondrial NADH and flavin adenine dinucleotide (FAD) autofluorescence in intact hearts. A mediating role of ATP-sensitive K⁺ (K_ATP) channel opening was investigated. NADH and FAD fluorescence was measured in the left ventricular wall of guinea pig isolated hearts assigned to five groups of eight animals each: hypothermia alone, hypothermia with ischemia, IPC with cold ischemia, 5-hydroxydecanoic acid (5-HD) alone, and 5-HD with IPC and cold ischemia. IPC consisted of two 5-min periods of warm global ischemia spaced 5 min apart and 15 min of reperfusion before 2 h of ischemia at 27°C and 2 h of warm reperfusion. The K_ATP channel inhibitor 5-HD was perfused from 5 min before until 5 min after IPC. IPC before 2 h of ischemia at 27°C led to better recovery of function and less tissue damage on reperfusion than did 27°C ischemia alone. These improvements were preceded by attenuated increases in NADH and decreases in FAD during cold ischemia and the reverse changes during warm reperfusion. 5-HD blocked each of these changes induced by IPC. This study indicates that IPC induces additive cardioprotection with mild hypothermic ischemia by improving mitochondrial bioenergetics during and after ischemia. Because effects of IPC on subsequent changes in NADH and FAD were inhibited by 5-HD, this suggests that mitochondrial K_ATP channel opening plays a substantial role in improving mitochondrial bioenergetics throughout mild hypothermic ischemia and reperfusion.

nicotinamide adenine dinucleotide; flavin adenine dinucleotide; ATP-sensitive potassium channels; 5-hydroxydecanoic acid

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