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).
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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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DOI: https://doi.org/10.1007/s11010-009-0283-2