Role of oxidative stress in ischemia-reperfusion-induced changes in Na+,K(+)-ATPase isoform expression in rat heart

The aim of this study was to assess whether depression of cardiac Na+,K(+)-ATPase activity during ischemia/reperfusion (I/R) is associated with alterations in Na+,K(+)-ATPase isoforms, and if oxidative stress participates in these I/R-induced changes. Na+,K(+)-ATPase alpha1, alpha2, alpha3, beta1, beta2, and beta3 isoform contents were measured in isolated rat hearts subjected to I/R (30 min of global ischemia followed by 60 min of reperfusion) in the presence or absence of superoxide dismutase plus catalase (SOD+CAT). Effects of oxidative stress on Na+,K(+)-ATPase isoforms were also examined by perfusing the hearts for 20 min with 300 microM hydrogen peroxide or 2 mM xanthine plus 0.03 U/ml xanthine oxidase (XXO). I/R significantly reduced the protein levels of all alpha and beta isoforms. Treatment of I/R hearts with SOD+CAT preserved the levels of alpha2, alpha3, beta1, beta2, and beta3 isoforms, but not that of the alpha1 isoform. Perfusion of hearts with hydrogen peroxide and XXO depressed all Na+,K(+)-ATPase alpha and beta isoforms, except for alpha1. These results indicate that the I/R-induced decrease in Na+,K(+)-ATPase may be due to changes in Na+,K(+)-ATPase isoform expression and that oxidative stress plays a role in this alteration. Antioxidant treatment attenuated the I/R-induced changes in expression of all isoforms except alpha1, which appears to be more resistant to oxidative stress.     

Captopril treatment improves the sarcoplasmic reticular Ca(2+) transport in heart failure due to myocardial infarction.

Although captopril, an angiotensin-converting enzyme (ACE) inhibitor, has been shown to exert a beneficial effect on cardiac function in heart failure, its effect on the status of sarcoplasmic reticulum (SR) Ca(2+) transport in the failing heart has not been examined previously. In order to determine whether captopril has a protective action on cardiac function, as well as cardiac SR Ca(2+)-pump activity and gene expression, a rat model of heart failure due to myocardial infarction was employed in this study. Sham operated and infarcted rats were given captopril (2 g/l) in drinking water; this treatment was started at either 3 or 21 days and was carried out until 8 weeks after the surgery. The untreated animals with myocardial infarction showed increased heart weight and elevated left ventricular end diastolic pressure, reduced rates of pressure development and pressure fall, as well as depressed SR Ca(2+) uptake and Ca(2+)-stimulated ATPase activities in comparison with the sham control group. These hemodynamic and biochemical changes in the failing hearts were prevented by treatment of the infarcted animals with captopril. Likewise, the observed reductions in the SR Ca(2+) pump and phospholamban protein contents, as well as in the mRNA levels for SR Ca(2+) pump ATPase and phospholamban, in the failing heart were attenuated by captopril treatment. These results suggest that heart failure is associated with a defect in the SR Ca(2+) handling and a depression in the gene expression of SR proteins; the beneficial effect of captopril in heart failure may be due to its ability to prevent remodeling of the cardiac SR membrane.     

Palmitoyl carnitine increases intracellular calcium in adult rat cardiomyocytes.

Earlier studies have demonstrated that palmitoyl carnitine (PC), a long chain acyl carnitine, accumulates in the ischemic myocardium. Although perfusion of hearts with PC is known to induce contractile dysfunction which resembles ischemic contracture, the mechanisms underlying this derangement are not clear. In this study, we examined the effect of exogenous PC on the intracellular concentration of calcium ([Ca(2+)](i)) in freshly isolated cardiomyocytes from adult rat hearts. The results showed that PC elevated [Ca(2+)](i)in a dose-dependent (5-20 microm) manner; 15 microm PC evoked a marked and reversible increase in [Ca(2+)](i)without having any significant action on cell viability. The PC (15 microm)-induced increase in [Ca(2+)](i)was slightly depressed but delayed in the absence of extracellular Ca(2+). Pre-incubation of cardiomyocytes with sarcolemmal (SL) l -type Ca(2+)-channel blockers, verapamil or diltiazem, and inhibitors of SL Na(+)-Ca(2+)exchanger such as Ni(2+)or amiloride, depressed the PC-evoked increase in [Ca(2+)](i)significantly. Ouabain, a Na(+)-K(+)ATPase inhibitor, and low concentrations of extracellular Na(+)enhanced the PC-induced increase in [Ca(2+)](i). Depletion of the sarcoplasmic reticulum (SR) Ca(2+)stores by low micromolar concentrations of ryanodine (a SR Ca(2+)-release channel activator) or by thapsigargin (a SR Ca(2+)-pump ATPase inhibitor) depressed the PC-mediated increase in [Ca(2+)](i). Combined blockade of the l -type Ca(2+)channel, Na(+)-Ca(2+)exchanger and the SR Ca(2+)-pump had an additive inhibitory effect on the PC response. These observations suggest that the PC-induced increase in [Ca(2+)](i)is dependent on both Ca(2+)-influx from the extracellular space and Ca(2+)-release from the SR stores. Thus, the accumulation of PC in the myocardium may be partly responsible for the occurrence of intracellular Ca(2+)overload in ischemic heart.     

Effects of some L-carnitine derivatives on heart membrane ATPases

In order to understand the role of carnitine metabolites in the genesis of cellular dysfunction and damage due to myocardial ischemia, the effects of 1–100 μM L-carnitine, acetylcarnitine, propionylcarnitine, and palmitoylcarnitine were investigated on rat heart sarcolemmal, sarcoplasmic reticular, and mitochondrial ATPase activities. Palmitoylcarnitine, unlike acetylcarnitine, propionylcarnitine and carnitine, produced marked inhibitory actions on sarcolemmal Na,K-ATPase and Ca²⁺-stimulated ATPase, as well as sarcoplasmic reticular Ca²⁺-stimulated ATPase activities; Na,K-ATPase was most sensitive. Although palmitoylcarnitine, unlike carnitine or its short-chain fatty-acid derivatives, also depressed sarcolemmal Ca²⁺ ATPase or Mg²⁺ ATPase, sarcoplasmic reticular Mg²⁺ ATPase, and mitochondrial Mg²⁺ ATPase, mitochondria were less sensitive in comparison to other organelles. Myofibrillar Ca²⁺-stimulated ATPase was slightly inhibited by very high concentrations of palmitoly-carnitine only. It is suggested that the observed depression of the sarcolemmal Na⁺-pump system by low concentrations of long-chain acyl derivatives of carnitine may contribute towards the pathogenesis of arrhythmias due to myocardial ischemia. Furthermore, the inhibition of Ca²⁺-pump mechanisms in the sarcolemmal and sarcoplasmic reticular membranes by relatively high concentrations of palmitoylcarnitine may result in the occurrence of intracellular Ca²⁺ overload and subsequent cell damage, as well as cardiac dysfunction due to myocardial ischemia.     

Role of cholesterol in cardiovascular dysfunction

BACKGROUND: Hypercholesterolemia, which is characterized by high levels of lipoprotein-containing cholesterol in plasma, is generally accepted as a major risk factor for the development of atherosclerosis and subsequent myocardial ischemia. The cardiovascular effects of elevated serum cholesterol are predominantly attributed to atherosclerotic lesions in coronary arteries; however, the role of cholesterol in causing heart dysfunction without the occurrence of atherosclerosis is not fully appreciated. OBSERVATIONS: Each type of biological membrane has a specific amount of cholesterol, which is required for proper functioning of the membrane-bound enzymes and cation transporters. High levels of cholesterol have been demonstrated to cause changes in membrane structure and function independent of atherosclerosis. Particularly, alterations in membrane cholesterol content have been shown to affect myocardial contractility, excitability and conduction properties. The activities of cardiac sarcolemmal enzymes such as Na+-K+ ATPase, Mg2+ ATPase and Ca2+ pump ATPase as well as Ca2+-dependent K+ channels and Na+-Ca2+ exchanger are altered as a consequence of changes in the membrane cholesterol content. Furthermore, high levels of cholesterol are known to cause endothelial dysfunction and smooth muscle abnormalities, which occur without visible atherosclerosis lesion development. On the other hand, indirect effects of cholesterol on the cardiovascular system are evident on its oxidative modification and subsequent development of visible atherosclerotic lesions and myocardial ischemia. CONCLUSIONS: Hypercholesterolemia has been shown to cause cardiovascular dysfunction due to direct action on membrane fluidity, enzyme activities and cation transporters in the endothelial cells, vascular smooth muscle cells and cardiomyocytes. On the other hand, development of atherosclerosis by high levels of cholesterol is associated with the oxidation products of cholesterol and is thus considered to affect the cardiovascular system indirectly as a consequence of ischemic heart disease.     

Insulin-induced enhancement of uptake of noradrenaline in atrial strips.

1 Addition of insulin to the organ bath increased the force of contraction of guinea-pig left atrial strips driven electrically at 1 Hz. 2 The positive inotropic response to insulin remained unaltered in atria depleted of catecholamine or when beta-adrenoceptors were blocked by addition of propranolol to the organ bath. 3 The response os isolated atria to noradrenaline was significantly reduced in the presence of insulin. 4 Insulin affected neither the calcium accumulating abilities of the heart sarcolemma, mitochondria or microsomes, nor the cyclic adenosine 3',5'-monophosphate (cyclic-AMP)-protein kinase-induced stimulation of microsomal calcium uptake. 5 Addition of insulin to the organ bath enhanced significantly the ability of the cardiac tissue to take up [3H]-noradrenaline as well as [3H]-metaraminol. The activities of monoamine oxidase and catechol-O-methyl transferase were not changed after addition of insulin to homogenates of the heart. 6 The ability of insulin to facilitate uptake of noradrenaline would be expected to cause a decrease in the amount of the amine reaching the receptors, thus leading to a diminished response to this amine. This may explain, at least in part, insulin-induced subsensitivity to noradrenaline. 7 This view is supported by the observation that after blockade of amine uptake by destruction of nerve terminals, insulin failed to reduce the positive inotropic response to noradrenaline.