Protein kinase C(epsilon) modulates apoptosis induced by beta -adrenergic stimulation in adult rat ventricular myocytes via extracellular signal-regulated kinase (ERK) activity.
Beta-adrenergic stimulation of ventricular myocytes has been shown to induce apoptosis; however, the cellular mechanisms involved in this pathway have not been completely characterized. This study examines the role of protein kinase C (PKC) in the signaling cascade that mediates beta-adrenergic stimulation-induced apoptosis. Stimulation of beta-adrenergic receptors using isoproterenol (ISO, 1-10 microm, 24 h) induced apoptosis in cultured adult rat ventricular myocytes (ARVM) in a dose-dependent manner. Treatment with ISO significantly resulted in the membrane translocation of PKC(epsilon), but not of PKC alpha or delta in ARVM. The activation of PKC(epsilon) by ISO was confirmed using an immune complex kinase assay. To address whether PKC(epsilon) is involved in the mechanism of ISO-induced apoptosis, we used the PKC(epsilon)-specific translocation inhibitor peptide, epsilonV1-2. Peptide epsilonV1-2 significantly blocked the translocation of PKC(epsilon), as well as the enzymatic action of PKC(epsilon), resulting from ISO stimulation. The inhibition of PKC(epsilon) attenuated ISO-induced apoptosis as measured by terminal deoxynucleotidyltransferase nick-end labeling (TUNEL) assay (18.2+/-3.8%v 49.0+/-2.4%P<0.05), while a PKC delta-specific peptide translocation inhibitor (delta V1-1) failed to do so (39.8+/-7.8%). In the presence of ISO, PKC(epsilon) inhibition by epsilonV1-2 was found to significantly enhance activity of ERK, but not that of Akt/PKB. Inhibition of ERK activation by PD 98059 (10-50 microm) attenuated the epsilonV1-2 peptide-mediated anti-apoptotic effect, thus suggesting that ERK activation is involved in this anti-apoptotic effect. Therefore, our results suggest that activation of PKC(epsilon) downstream of beta-adrenergic stimulation promotes apoptosis largely via inhibition of an ERK activation-dependent anti-apoptotic effect.
Shizukuda
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Program in Cardiovascular Sciences, Section of Cardiology, Department of Medicine, University of Illinois at Chicago, 60612, USA. shizukud@uic.edu
Buttrick
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PM
D000318:Adrenergic beta-Agonists / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D006352:Heart Ventricles / Q000201:enzymology*; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D007527:Isoenzymes / Q000378:metabolism* / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism* / Q000502:physiology*; D051744:Protein Kinase C-epsilon; D021381:Protein Transport; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction
D000318:Adrenergic beta-Agonists; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007527:Isoenzymes; D010455:Peptides; C497257:Prkce protein, rat; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D020928:Mitogen-Activated Protein Kinases; D007545:Isoproterenol; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one