Role of protein kinase C in mediating effects of hydrogen peroxide in guinea-pig ventricular myocytes.
The present study examined the effects of hydrogen peroxide (H2O2) on intracellular calcium transients and unloaded cell shortening in the presence of the protein kinase C (PKC) inhibitors 1-(5-isoquinolinesulfonyl-2-methylpiperazine (H7) or chelerythrine chloride (CHC) or the PKC activator phorbol 12-myristate 13-acetate (PMA). Calcium transient amplitudes and cell shortening were measured simultaneously in single, enzymatically dissociated ventricular myocytes loaded with fura2-AM. Exposure of myocytes to H2O2, 25 microM or 75 microM, for 15 min caused a time- and concentration-dependent increase in calcium transient amplitude, cell shortening and the diastolic 340/380 fluorescence ratio. Significant increases in calcium transient amplitude were observed from 7 to 15 min of superfusion with 25 microM H2O2 and the transient amplitude remained elevated throughout the 10 min washout period. In the presence of 75 microM H2O2, transient amplitude was elevated following 2 min and remained elevated for the remainder of the experiment. Significant increases in cell shortening were also observed from 7 to 15 min in the presence of either 25 or 75 microM H2O2. This effect was reversed upon washout of the lower concentration of H2O2 but persisted during the initial 5 min of washout at the higher concentration. The diastolic 340/380 fluorescence ratio was unaltered in the presence of 25 microM of H2O2, however this parameter was significantly increased from 7 to 15 min following exposure to 75 microM H2O2 and remained elevated throughout the washout period. The H2O2-induced increases in calcium transient amplitude and cell shortening were significantly attenuated in myocytes which were pretreated with either H7 or CHC.(ABSTRACT TRUNCATED AT 250 WORDS)
Ward
C A
CA
Department of Pharmacology and Toxicology, University of Western Ontario, London, Canada.
Moffat
M P
MP
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002118:Calcium / Q000378:metabolism*; D048429:Cell Size / Q000187:drug effects; D004789:Enzyme Activation; D006168:Guinea Pigs; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism; D006706:Homeostasis / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D066298:In Vitro Techniques; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D015425:Myocardial Reperfusion; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010617:Phenanthridines / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors
D000470:Alkaloids; D053119:Benzophenanthridines; D007546:Isoquinolines; D010617:Phenanthridines; D010879:Piperazines; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D006861:Hydrogen Peroxide; C016299:chelerythrine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium