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Cardioprotection initiated by reactive oxygen species is dependent on activation of PKCε.
American Journal of Physiology - Heart and Circulatory Physiology
To examine whether cardioprotection initiated by reactive oxygen species (ROS) is dependent on protein kinase Cε (PKCε), isolated buffer-perfused mouse hearts were randomized to four groups: 1) antimycin A (AA) (0.1 μg/ml) for 3 min followed by 10 min washout and then 30 min global ischemia (I) and 2 h reperfusion (R); 2) controls of I/R alone; 3) AA bracketed with 13 min of N-2-mercaptopropionylglycine (MPG) followed by I/R; and 4) MPG (200 μM) alone, followed by I/R. Isolated adult rat ventricular myocytes (ARVM) were exposed to AA (0.1 μg/ml), and lucigenin was used to measure ROS production. Murine hearts and ARVM were exposed to AA (0.1 μg/ml) with or without MPG, and PKCε translocation was measured by cell fractionation and subsequent Western blot analysis. Finally, the dependence of AA protection on PKCε was determined by the use of knockout mice (-/-) lacking PKCε. AA exposure caused ROS production, which was abolished by the mitochondrial uncoupler mesoxalonitrile 4-trifluoromethoxyphenylhydrazone. In addition, AA significantly reduced the percent infarction-left ventricular volume compared with control I/R (26 ± 4 vs. 43 ± 2%; P < 0.05). Bracketing AA with MPG caused a loss of protection (52 ± 7 vs. 26 ± 4%; P < 0.05). AA caused PKCε translocation only in the absence of MPG, and protection was lost on the pkcε background (38 ± 3 vs. 15 ± 4%; P < 0.001). AA causes ROS production, on which protection and PKCε translocation depend. In addition, protection is absent in PKCε null hearts. Our results imply that, in common with ischemic preconditioning, PKCε is crucial to ROS-mediated protection. Copyright © 2006 the American Physiological Society.
|Title:||Cardioprotection initiated by reactive oxygen species is dependent on activation of PKCε|
|UCL classification:||UCL > School of Life and Medical Sciences > Faculty of Medical Sciences > Medicine (Division of)|
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