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Am J Physiol Cell Physiol 294: C169-C177, 2008. First published November 7, 2007; doi:10.1152/ajpcell.00282.2007
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RECEPTORS AND SIGNAL TRANSDUCTION

H2S preconditioning-induced PKC activation regulates intracellular calcium handling in rat cardiomyocytes

Ting-Ting Pan, Kay Li Neo, Li-Fang Hu, Qian Chen Yong, and Jin-Song Bian

Cardiovascular Biology Research Group, Department of Pharmacology, National University of Singapore, Singapore

Submitted 7 August 2007 ; accepted in final form 15 November 2007

The present study was aimed to investigate the regulatory effect of protein kinase C (PKC) on intracellular Ca2+ handling in hydrogen sulfide (H2S)-preconditioned cardiomyocytes and its consequent effects on ischemia challenge. Immunoblot analysis was used to assess PKC isoform translocation in the rat cardiomyocytes 20 h after NaHS (an H2S donor, 10–4 M) preconditioning (SP, 30 min). Intracellular Ca2+ was measured with a spectrofluorometric method using fura-2 ratio as an indicator. Cell length was compared before and after ischemia-reperfusion insults to indicate the extent of hypercontracture. SP motivated translocation of PKC{alpha}, PKC{varepsilon}, and PKC{delta} to membrane fraction but only translocation of PKC{varepsilon} and PKC{delta} was abolished by an ATP-sensitive potassium channel blocker glibenclamide. It was also found that SP significantly accelerated the decay of both electrically and caffeine-induced intracellular [Ca2+] transients, which were reversed by a selective PKC inhibitor chelerythrine. These data suggest that SP facilitated Ca2+ removal via both accelerating uptake of Ca2+ into sarcoplasmic reticulum and enhancing Ca2+ extrusion through Na+/Ca2+ exchanger in a PKC-dependent manner. Furthermore, blockade of PKC also attenuated the protective effects of SP against Ca2+ overload during ischemia and against myocyte hypercontracture at the onset of reperfusion. We demonstrate for the first time that SP activates PKC{alpha}, PKC{varepsilon}, and PKC{delta} in cardiomyocytes via different signaling mechanisms. Such PKC activation, in turn, protects the heart against ischemia-reperfusion insults at least partly by ameliorating intracellular Ca2+ handling.

protein kinase C isoforms; ischemia and reperfusion; cardioprotection; ATP-sensitive potassium channel



Address for reprint requests and other correspondence: J.-S. Bian, Cardiovascular Biology Research Group, Dept. of Pharmacology, National Univ. of Singapore, Singapore, 117597 (e-mail: phcbjs{at}nus.edu.sg)




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Q. C. Yong, S. W. Lee, C. S. Foo, K. L. Neo, X. Chen, and J.-S. Bian
Endogenous hydrogen sulphide mediates the cardioprotection induced by ischemic postconditioning
Am J Physiol Heart Circ Physiol, September 1, 2008; 295(3): H1330 - H1340.
[Abstract] [Full Text] [PDF]




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