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Br J Pharmacol. 2010 Nov;161(5):1172-85. doi: 10.1111/j.1476-5381.2010.00960.x.

Pharmacological preconditioning by diazoxide downregulates cardiac L-type Ca(2+) channels.

Author information

1
Departamento de Farmacología, Centro de Investigación y de Estudios Avanzados del I.P.N. México, México.

Abstract

BACKGROUND AND PURPOSE:

Pharmacological preconditioning (PPC) with mitochondrial ATP-sensitive K(+) (mitoK(ATP) ) channel openers such as diazoxide, leads to cardioprotection against ischaemia. However, effects on Ca(2+) homeostasis during PPC, particularly changes in Ca(2+) channel activity, are poorly understood. We investigated the effects of PPC on cardiac L-type Ca(2+) channels.

EXPERIMENTAL APPROACH:

PPC was induced in isolated hearts and enzymatically dissociated cardiomyocytes from adult rats by preincubation with diazoxide. We measured reactive oxygen species (ROS) production and Ca(2+) signals associated with action potentials using fluorescent probes, and L-type currents using a whole-cell patch-clamp technique. Levels of the α(1c) subunit of L-type channels in the cellular membrane were measured by Western blot.

KEY RESULTS:

PPC was accompanied by a 50% reduction in α(1c) subunit levels, and by a reversible fall in L-type current amplitude and Ca(2+) transients. These effects were prevented by the ROS scavenger N-acetyl-L-cysteine (NAC), or by the mitoK(ATP) channel blocker 5-hydroxydecanoate (5-HD). PPC significantly reduced infarct size, an effect blocked by NAC and 5-HD. Nifedipine also conferred protection against infarction when applied during the reperfusion period. Downregulation of the α(1c) subunit and Ca(2+) channel function were prevented in part by the protease inhibitor leupeptin.

CONCLUSIONS AND IMPLICATIONS:

PPC downregulated the α(1c) subunit, possibly through ROS. Downregulation involved increased degradation of the Ca(2+) channel, which in turn reduced Ca(2+) influx, which may attenuate Ca(2+) overload during reperfusion.

PMID:
20636393
PMCID:
PMC2998696
DOI:
10.1111/j.1476-5381.2010.00960.x
[Indexed for MEDLINE]
Free PMC Article

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