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Diazoxide-induced respiratory inhibition – a putative mitochondrial KATP channel independent mechanism of pharmacological preconditioning

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Abstract

The ischemic preconditioning biological phenomenon has been explored to identify putative pharmacologic agents to mimic this cytoprotective program against cellular ischemic injury. Diazoxide administration confers this cytoprotection, however, whether this is via direct activation of the putative mitochondrial KATP (mKATP) channel which was originally proposed has been questioned. Here, we present data supporting an alternate hypothesis evoking mitochondrial respiratory inhibition rather than mKATP channel activation, as a mediating event in the diazoxide-activated cytoprotective program. Mitochondrial respiration and reactive oxygen species (ROS) production was measured in digitonin-permeabilized C2C12 myotubes, allowing for the modulation of mKATP conductance by changing the potassium concentration of the medium (0–130 mM). Diazoxide dose-dependently attenuated succinate-supported respiration, an effect that was independent of mKATP channel conductance. Similarly, 5-hydroxydecanoate (5-HD), a putative mKATP channel blocker, released diazoxide-induced respiratory inhibition independently of potassium concentration. Since diazoxide-induced cytoprotection and respiratory inhibition are both integrally linked to ROS generation we repeated above experiments following ROS generation using DCF fluorescence. Cytoprotective doses of diazoxide increased ROS generation independently of potassium concentration and 5-HD inhibited ROS production under the same conditions. Collectively these data support the hypothesis that diazoxide-mediated cytoprotection is independent of the conductance of the mKATP channel and rather implicate mitochondrial respiratory inhibition-triggered ROS signaling.

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Authors and Affiliations

  1. Hatter Institute for Cardiology Research, MRC Inter-University Cape Heart Group, University of Cape Town Medical School, Cape Town, South Africa

    Jan Minners, Lydia Lacerda, Lionel H. Opie, Christopher J. McLeod & Michael N. Sack

  2. Hatter Institute for Cardiovascular Studies, University College Hospitals and Medical School, London, United Kingdom

    Derek M. Yellon

  3. NIH/NHLBI, Building 10-CRC, 5-3150, Bethesda, MD, 20892-1485, USA

    Michael N. Sack

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  1. Jan Minners
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  2. Lydia Lacerda
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  3. Derek M. Yellon
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  4. Lionel H. Opie
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  5. Christopher J. McLeod
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Correspondence to Michael N. Sack.

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Minners, J., Lacerda, L., Yellon, D.M. et al. Diazoxide-induced respiratory inhibition – a putative mitochondrial KATP channel independent mechanism of pharmacological preconditioning. Mol Cell Biochem 294, 11–18 (2007). https://doi.org/10.1007/s11010-005-9066-6

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  • DOI: https://doi.org/10.1007/s11010-005-9066-6

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