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Opening of the mitoKATP channel and decoupling of mitochondrial complex II and III contribute to the suppression of myocardial reperfusion hyperoxygenation

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Abstract

Diazoxide, a mitochondrial ATP-sensitive potassium (mitoKATP) channel opener, protects the heart from ischemia–reperfusion injury. Diazoxide also inhibits mitochondrial complex II-dependent respiration in addition to its preconditioning effect. However, there are no prior studies of the role of diazoxide on post-ischemic myocardial oxygenation. In the current study, we determined the effect of diazoxide on the suppression of post-ischemic myocardial tissue hyperoxygenation in vivo, superoxide (O2 −•) generation in isolated mitochondria, and impairment of the interaction between complex II and complex III in purified mitochondrial proteins. It was observed that diazoxide totally suppressed the post-ischemic myocardial hyperoxygenation. With succinate but not glutamate/malate as the substrate, diazoxide significantly increased ubisemiquinone-dependent O2 −• generation, which was not blocked by 5-HD and glibenclamide. Using a model system, the super complex of succinate-cytochrome c reductase (SCR) hosting complex II and complex III, we also observed that diazoxide impaired complex II and its interaction with complex III with no effect on complex III. UV–visible spectral analysis revealed that diazoxide decreased succinate-mediated ferricytochrome b reduction in SCR. In conclusion, our results demonstrated that diazoxide suppressed the in vivo post-ischemic myocardial hyperoxygenation through opening the mitoKATP channel and ubisemiquinone-dependent O2 −• generation via inhibiting mitochondrial complex II-dependent respiration.

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Abbreviations

DCPIP:

Dichlorophenolindophenol

DEPMPO:

5-(Diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide

DMPO:

5,5-Dimethyl-1-pyrroline N-oxide

DTPA:

Diethylenetriaminepentaacetic acid

EPR:

Electron paramagnetic resonance

FCCP:

Carbonyl cyanide-p-tri-fluromethoxyphenyl-hydrazone

5-HD:

5-hydroxydecanoate

H2DCF:

2′,7′-dichlorodihydrofluorescein

IPC:

Ischemic preconditioning

mitoKATP :

Mitochondrial ATP-sensitive potassium channel

3-NPA:

3-Nitropropionic acid

O2 −• :

Superoxide

•OH:

Hydroxyl radical

\( P_{{{\text{O}}_{ 2} }} \) :

Tissue oxygen tension

PPC:

Pharmacological preconditioning

Q:

Ubiquinone

Q−• :

Ubisemiquinone

Q2 :

Ubiquinone-2

QCR:

Ubiquinol cytochrome c reductase or complex III

QH2 :

Ubiquinol

QO −• :

Ubisemiquinone at the QO site

ROS:

Reactive oxygen species

SCR:

Succinate-cytochrome c reductase or super complex containing complex II and complex III

SMP:

Sub-mitochondrial particles

SOD:

Superoxide dismutase

SQR:

Succinate ubiquinone reductase or complex II

TEA+ :

Tetraethyl ammonium

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Acknowledgments

This work was supported by American Heart Association Grant 0435299N and National Heart, Lung, and Blood Institute Grants HL-081630 (He) and HL-083237 (Chen). The LiPc was made with the support of PO1 EB2180 (Swartz).

Author information

Author notes

  1. Bin Liu

    Present address: Shantou University Medical College, Shantou, 515041, China

Authors and Affiliations

  1. The Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, The Ohio State University College of Medicine, Columbus, OH, 43210, USA

    Bin Liu

  2. Key Laboratory of Organ Transplantation, Ministry of Education Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Xuehai Zhu

  3. Davis Heart and Lung Research Institute, The Ohio State University College of Medicine, Columbus, OH, 43210, USA

    Chwen-Lih Chen & Yeong-Renn Chen

  4. Division of Pharmacology, College of Pharmacy, The Ohio State University, Columbus, OH, USA

    Keli Hu

  5. The EPR Center for the Study of Viable Systems, Dartmouth Medical School, Hanover, NH, USA

    Harold M. Swartz

  6. The Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine, Department of Internal Medicine, The Ohio State University College of Medicine, 460 West 12th Avenue, 0388/BRT, Columbus, OH, 43210, USA

    Guanglong He

  7. Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine, Department of Internal Medicine, The Ohio State University College of Medicine, 460 West 12th Avenue, 0388/BRT, Columbus, OH, 43210, USA

    Guanglong He

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  1. Bin Liu
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Correspondence to Guanglong He.

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Liu, B., Zhu, X., Chen, CL. et al. Opening of the mitoKATP channel and decoupling of mitochondrial complex II and III contribute to the suppression of myocardial reperfusion hyperoxygenation. Mol Cell Biochem 337, 25–38 (2010). https://doi.org/10.1007/s11010-009-0283-2

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