Oxidoreductase regulation of Kv currents in rat ventricle.
Oxidative stress contributes to the arrhythmogenic substrate created by myocardial ischemia-reperfusion partly through a shift in cell redox state, a key modulator of protein function. The activity of many oxidation-sensitive proteins is controlled by oxidoreductase systems that regulate the redox state of cysteine thiol groups, but the impact of these systems on ion channel function is not well defined. Thus, we examined the roles of the thioredoxin and glutaredoxin systems in controlling K(+) channels in the ventricle. An oxidative shift in redox state was elicited in isolated rat ventricular myocytes by brief exposure to diamide, a thiol-specific, membrane-permeable oxidant. Voltage-clamp studies showed that diamide decreased peak outward K(+) current (I(peak)) evoked by depolarizing test pulses by 41% (+60 mV; p<0.05) while steady-state outward current (I(ss)) measured at the end of the test pulse was decreased by 45% (p<0.05). These electrophysiological effects were not prevented by protein kinase C blockers, but the tyrosine kinase inhibitors genistein or lavendustin A blocked the suppression of both K(+) currents by diamide. Moreover, inhibition of I(peak) and I(ss) by diamide was reversed by dichloroacetate and an insulin-mimetic. The effect of dichloroacetate to normalize I(peak) after diamide was blocked by the thioredoxin system inhibitors auranofin or 13-cis-retinoic acid, but I(ss) was not affected by either compound. A pan-specific inhibitor of glutaredoxin and thioredoxin systems, 1,3-bis-(2-chloroethyl)-1-nitrosourea, also blocked the dichloroacetate effect on I(peak) but only partially inhibited the recovery of I(ss). These data suggest that acute regulation of cardiac K(+) channels by oxidoreductase systems is mediated by redox-sensitive tyrosine kinase/phosphatase pathways. The pathways controlling I(peak) channels are targets of the thioredoxin system whereas those regulating I(ss) channels are likely controlled by the glutaredoxin system. Thus, cardiac oxidoreductase systems may be important regulators of ion channels affected by pathogenic oxidative stress.
Liang
Huixu
H
Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, USA.
Li
Xun
X
Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, USA; Department of Cardiology, The First Affiliated Hospital, Soochow University, Suzhou, Jiangsu, P.R. China.
Li
Shumin
S
Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, USA.
Zheng
Ming-Qi
MQ
Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, USA.
Rozanski
George J
GJ
Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, USA; Center for Redox Biology, University of Nebraska-Lincoln, Lincoln, Nebraska, USA. Electronic address: grozansk@unmc.edu.
D000818:Animals; D003999:Dichloroacetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D054477:Glutaredoxins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000201:enzymology*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D018408:Patch-Clamp Techniques; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013879:Thioredoxins / Q000037:antagonists & inhibitors / Q000378:metabolism*
D004791:Enzyme Inhibitors; D054477:Glutaredoxins; D016877:Oxidants; D024642:Potassium Channels, Voltage-Gated; D013879:Thioredoxins; D003999:Dichloroacetic Acid; D011505:Protein-Tyrosine Kinases; D010744:Phosphoric Monoester Hydrolases