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Am J Physiol Heart Circ Physiol 280: H280-H293, 2001;
0363-6135/01 $5.00
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Vol. 280, Issue 1, H280-H293, January 2001

Changes in [Na+]i, compartmental [Ca2+], and NADH with dysfunction after global ischemia in intact hearts

Srinivasan G. Varadarajan, Jianzhong An, Enis Novalija, Steven C. Smart, and David F. Stowe

Anesthesiology Research Laboratory, Departments of Medicine (Cardiovascular Diseases), Anesthesiology, and Physiology, Medical College of Wisconsin and Cardiovascular Research Center, Milwaukee 53226, and Research Service, Veterans Affairs Medical Center, Milwaukee, Wisconsin 53295

We measured the effects of global ischemia and reperfusion on intracellular Na+, NADH, cytosolic and mitochondrial (subscript mito) Ca2+, relaxation, metabolism, contractility, and Ca2+ sensitivity in the intact heart. Langendorff-prepared guinea pig hearts were crystalloid perfused, and the left ventricular (LV) pressure (LVP), first derivative of LVP (LV dP/dt), coronary flow, and O2 extraction and consumption were measured before, during, and after 30-min global ischemia and 60-min reperfusion. Ca2+, Na+, and NADH were measured by luminescence spectrophotometry at the LV free wall using indo 1 and sodium benzofuran isophthalate, respectively, after subtracting changes in tissue autofluorescence (NADH). Mitochondrial Ca2+ was assessed by quenching cytosolic indo 1 with MnCl2. Mechanical responses to changes in cytosolic-systolic (subscript sys), diastolic (subscript dia), and mitochondrial Ca2+ were tested over a range of extracellular [Ca2+] before and after ischemia-reperfusion. Both [Ca2+]sys and [Ca2+]dia doubled at 1-min reperfusion but returned to preischemia values within 10 min, whereas [Ca2+]mito was elevated over 60-min reperfusion. Reperfusion dissociated [Ca2+]dia and [Ca2+]sys from contractile function as LVPsys-dia and the rise in LV dP/dt (LV dP/dtmax) were depressed by one-third and the fall in LV dP/dt (LV dP/dtmin) was depressed by one-half at 30-min reperfusion, whereas LVPdia remained markedly elevated. [Ca2+]sys-dia sensitivity at 100% LV dP/dtmax was not altered after reperfusion, but [Ca2+]dia at 100% LV dP/dtmin and [Ca2+]mito at 100% LV dP/dtmax were markedly shifted right on reperfusion (ED50 +36 and +125 nM [Ca2+], respectively) with no change in slope. NADH doubled during ischemia but returned to normal on initial reperfusion. The intracellular [Na+] ([Na+]i) increased minimally during ischemia but doubled on reperfusion and remained elevated at 60-min reperfusion. Thus Na+ and Ca2+ temporally accumulate during initial reperfusion, and cytosolic Ca2+ returns toward normal, whereas [Na+]i and [Ca2+]mito remain elevated on later reperfusion. Na+ loading likely contributes to Ca2+ overload and contractile dysfunction during reperfusion.

cardiac injury; contractility and relaxation; cytosolic Ca2+; mitochondrial Ca2+; myocardium; intracellular sodium concentration


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