Protein kinase C-epsilon is a trigger of delayed cardioprotection against myocardial ischemia of kappa-opioid receptor stimulation in rat ventricular myocytes.
Kappa-opioid receptor (OR) stimulation with a selective agonist, U50,488H (U50), known to mediate the delayed cardioprotection of metabolic inhibition preconditioning (MIP) against cell injury/death in rat ventricular myocytes, has been shown to act via protein kinase C (PKC). We attempted to identify the PKC isoform(s) that is activated, thus triggering delayed cardioprotection of MIP and pretreatment with 10 microM U50 (U50 pretreatment, UP). Release of lactate dehydrogenase and exclusion of trypan blue by isolated rat ventricular myocytes were used as indices of cell injury and death, respectively. Both MIP and UP induced translocation of PKC-epsilon, but not other PKC isoforms, -alpha and -delta, from cytosolic to membrane fractions. This was accompanied by reductions in cell injury/death induced by lethal simulated ischemia. The effects of MIP and UP were attenuated and abolished by 1 microM nor-binaltorphimine, a selective kappa-OR antagonist, administered before and during preconditioning/pretreatment, respectively. The effects were mimicked by 10 nM phorbol-12-myristate-13-acetate, a PKC activator, but attenuated by 5 microM chelerythrine, a PKC inhibitor. More importantly, 0.1 microM epsilonV1-2, a selective PKC-epsilon inhibitor administered before and during MIP/UP, also attenuated the effects of both treatments on cell injury/death and translocation of PKC-epsilon. On the other hand, 5 microM rottlerin, a selective PKC-delta inhibitor, did not alter the effects of either treatment on injury/death. The results indicate that both MIP and UP activate PKC-epsilon, leading to delayed cardioprotection in rat ventricular myocytes.
Wang
G Y
GY
Department of Physiology and Institute of Cardiovascular Sciences and Medicine, Faculty of Medicine, The University of Hong Kong, SAR, China.
Zhou
J J
JJ
Shan
J
J
Wong
T M
TM
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology; D015153:Blotting, Western; D004396:Coloring Agents; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006352:Heart Ventricles / Q000166:cytology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D017202:Myocardial Ischemia / Q000235:genetics / Q000503:physiopathology / Q000517:prevention & control*; D009206:Myocardium / Q000166:cytology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D051744:Protein Kinase C-epsilon; D051381:Rats; D017207:Rats, Sprague-Dawley; D017465:Receptors, Opioid, delta / Q000037:antagonists & inhibitors; D017473:Receptors, Opioid, kappa / Q000819:agonists* / Q000235:genetics; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014178:Translocation, Genetic / Q000187:drug effects; D014343:Trypan Blue
D000098:Acetophenones; D001578:Benzopyrans; D004396:Coloring Agents; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D017465:Receptors, Opioid, delta; D017473:Receptors, Opioid, kappa; C085746:rottlerin; D007770:L-Lactate Dehydrogenase; C497257:Prkce protein, rat; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D014343:Trypan Blue; D013755:Tetradecanoylphorbol Acetate