Role of opioid delta1 receptors, mitochondrial K(ATP) channels, and protein kinase C during cardiocyte apoptosis.
Opioids attenuate cardiac injury after ischemia and reperfusion. We wanted to determine whether the protection of opioids is mediated by blocking cardiocyte apoptosis, and if so, to describe the role of opioid delta1 receptors and protein kinase C (PKC) in this effect. Chick embryonic cardiomyocytes were subjected to 12 h of simulated ischemia and then 12 h of re-oxygenation, which resulted in 54+/-3% (n=6) of cell apoptosis (n=6) as measured by flow cytometry. This result was consistent with DNA laddering and TUNEL assay. Preconditioning, elicited with three cycles of 1 min of simulated ischemia separated by 5 min of reoxygenation before prolonged simulated ischemia, reduced apoptosis (36+/-4%, n=6*). Pretreatment with BNTX (0.1 micromol/l), a selective opioid delta1 receptor blocker, abolished the effects of preconditioning (57+/-5%, n=6). The selective opioid delta receptor agonist BW373U86 (20 pmol/l) also attenuated apoptosis (39+/-3%, n=6* v control). These effects were abolished by 5-hydroxydecanoate (100 microm), a selective mitochondrial K(ATP) channel blocker (50+/-5%, n=6) and by Go-6976 (0.1 micromol/l), a specific PKC inhibitor. Both preconditioning and BW373U86 activated the PKC delta isoform of particulate fraction before simulated ischemia without effect on total and cytosolic fractions. Stimulation of opioid delta1 receptors activates mitochondrial K(ATP) channels and the PKC delta isoform in cultured ventricular myocytes. This is one important signal transduction pathway through which ischemic preconditioning blocks apoptosis and preserves cardiac function.
Liu
H
H
Department of Anesthesiology, The University of North Carolina, Chapel Hill, NC 27599-7010, USA.
Zhang
H Y
HY
McPherson
B C
BC
Baman
T
T
Roth
S
S
Shao
Z
Z
Zhu
X
X
Yao
Z
Z
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D001549:Benzamides / Q000494:pharmacology; D002478:Cells, Cultured; D002642:Chick Embryo; D004247:DNA / Q000378:metabolism; D005434:Flow Cytometry; D020287:In Situ Nick-End Labeling; D007527:Isoenzymes / Q000502:physiology*; D008928:Mitochondria / Q000378:metabolism*; D017202:Myocardial Ischemia; D009206:Myocardium / Q000166:cytology* / Q000473:pathology*; D010100:Oxygen / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D015221:Potassium Channels / Q000502:physiology*; D020033:Protein Isoforms; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D051745:Protein Kinase C-delta; D017465:Receptors, Opioid, delta / Q000037:antagonists & inhibitors / Q000502:physiology*; D015398:Signal Transduction; D013997:Time Factors
D001549:Benzamides; D007527:Isoenzymes; D010879:Piperazines; D015221:Potassium Channels; D020033:Protein Isoforms; D017465:Receptors, Opioid, delta; C083877:BW 373U86; D000255:Adenosine Triphosphate; D004247:DNA; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D010100:Oxygen