Monensin-Induced Increase in Intracellular Na+ Induces Changes in Na+ and Ca2+ Currents and Regulates Na+-K+ and Na+-Ca2+ Transport in Cardiomyocytes.

BACKGROUND/AIMS: Monensin, an Na ionophore, increases intracellular Na ([Na]i). Alteration of [Na]i influences ion transport through the sarcolemmal membrane. So far, the effects of monensin on ventricular myocytes have not been examined in detail. The main objective of this study was to elucidate the mechanism via which monensin-evoked increases in [Na]i affect the membrane potential and currents in ventricular myocytes of guinea pigs. METHODS: Membrane potentials and currents were measured using the whole-cell patch-clamp technique in single myocytes. The concentration of intracellular Ca ([Ca]i) was evaluated by measuring fluorescence intensity of Fluo-4. RESULTS: Monensin (10-5M) shortened the action potential duration (APD) and reduced the amplitude of the plateau phase. In addition, monensin decreased the sodium current (INa) and shifted the inactivation curve to the hyperpolarized direction. Moreover, it decreased the L-type calcium current (ICa). However, this effect was attenuated by increasing the buffering capacity of [Ca]i. The Na-Ca exchange current (INa-Ca) was activated particularly in the reverse mode. Na-K pump current (INa-K) was also activated. Notably, the inward rectifying K current (IK1) was not affected, and the change in the delayed outward K current (IK) was not evident. CONCLUSION: These results suggest that the monensin-induced shortened APD and reduced amplitude of the plateau phase are primarily due to the decrease in the ICa, the activation of the reverse mode of INa-Ca, and the increased INa-K, and second due to the decreased INa. The IK and the IK1 may not be associated with the abovementioned changes induced by monensin. The elevation of [Na]i can exert multiple influences on electrophysiological phenomena in cardiac myocytes.

Cardio-protection by Ginkgo biloba extract 50 in rats with acute myocardial infarction is related to Na⁺-Ca²âº exchanger.

Ginkgo biloba has been used for medical purposes for centuries in traditional Chinese medicine. Ginkgo biloba extract 50 (GBE50) is a new standardized GBE product that matches the standardized German product as EGb761. This paper is aimed at studying the cardio-protection effects of GBE50 Salvia miltiorrhiza on myocardial function, area at risk, myocardial ultra-structure, and expression of calcium handling proteins in rat ischemic myocardium. Myocardium ischemia was induced by the left anterior descending (LAD) coronary artery occlusion and myocardial function was recorded by a transducer advanced into the left ventricle on a computer system. In vitro myocardial infarction was measured by 2,3,5-triphenyltetrazolium chloride (TTC) and Evans blue staining of heart sections. Morphological change was evaluated by electric microscopy and Western blotting was used for protein expression. Hemodynamic experiments in vivo showed that postischemic cardiac contractile function was reduced in ischemic rats. Salvia miltiorrhiza (7.5 g/kg/d×7) and Ginkgo biloba extract 50 (GBE50) (100 mg/kg/d×7) improved post-schemic cardiac diastolic dysfunction while not affecting the systolic function. In hearts of GBE50 group and Salvia miltiorrhiza (SM) group, the area at risk was significantly reduced and myocardial structure was better-preserved. Moreover, Na⁺-Ca²âº exchanger (NCX) expression increase and sarcoplasmic reticulum Ca²âº-ATPase 2 (SERCA2), LTCC, and ryanodine receptor 2 (RyR2) expression decreases were smaller than those in ischemia group. There was a significant difference between the GBE50 and ischemia group in NCX expression. GBE50 could improve recovery in contractile function and prevent myocardium from ischemia damage, which may be caused by attenuating the abnormal expression of NCX.

Overexpression of HSP-70 attenuates increases in [Ca2+]i and protects human epidermoid A-431 cells after chemical hypoxia.

This laboratory previously reported that thermotolerance diminishes the NaCN-induced increase in intracellular free calcium concentrations ([Ca2+]i) in human epidermoid A-431 cells and that blocking this increase protects the cells from NaCN toxicity. In this study, we report that cell viability after exposure to NaCN (10 mM, 1 h) is enhanced by the overexpression of HSP-70 resulting from heat shock (45 degrees C, 10 min), treatment with a protein kinase C activator phorbol 12 myristate 13-acetate (PMA; 1 microM, 4 h), or HSP-70 cDNA transfection. Because the toxicity of NaCN is mediated by increases in [Ca2+]i, we sought to determine whether the overexpression of HSP-70 might protect the cells by altering the [Ca2+]i response induced by NaCN. Basal [Ca2+]i in vector-, HSF1 cDNA-, and HSP-70 cDNA-transfected cells was 114 +/- 11 (n = 11), 95 +/- 5 (n = 6), and 151 +/- 11 (n = 15) nM, respectively, suggesting that HSP-70 metabolism is associated with maintenance of resting [Ca2+]i. Removal of external Ca2+ reduced the resting [Ca2+]i in all of these cells. With external Ca2+ reduced the resting [Ca2+]i by 97 +/- 21% in vector-transfected cells and 111 +/- 5% in HSF1 vector-transfected cells but by only 27 +/- 8% in HSP-70 cDNA-transfected cells. Heat shock or PMA treatment of vector- or HSF1 cDNA-transfected cells to induce HSP-70 also attenuated the NaCN-induced increase in [Ca2+]i, perhaps because of a decrease in Vmax for the uptake of external Ca2+. Removal of external Ca2+ or treatment with inhibitors of Na+/Ca2+ exchangers eliminated the NaCN-induced increase in [Ca2+]i in HSP-70 cDNA-transfected cells, but ryanodine treatment did not. HSP-70 cDNA transfection also reduced Ca2+ mobilization stimulated by various Ca(2+)-mobilizing agents. The results suggest that HSP-70 overexpression protects cells from NaCN cytotoxicity, perhaps by attenuating the [Ca2+]i response.