Downregulation of protein kinase C inhibits activation of mitochondrial K(ATP) channels by diazoxide.

Abstract

BACKGROUND:

The mitochondrial K(ATP) (mitoK(ATP)) channel has been shown to confer short- and long-term cardioprotection against prolonged ischemia via protein kinase C (PKC) signaling pathways. However, the exact association between PKC or its isoforms and mitoK(ATP) channels has not yet been clarified. The present study tested the hypothesis that the activity and translocation of PKC to the mitochondria are important for cardiac protection elicited by mitoK(ATP) channels. Methods and Results-PKC was downregulated by prolonged (24-hour) treatment with phorbol 12-myristate 13-acetate (4 microgram/kg body weight) before subsequent experiments in rats. Langendorff-perfused rat hearts were subjected to 40 minutes of ischemia followed by 30 minutes of reperfusion. Effects of PKC downregulation on the activation of mitoK(ATP) channels and other interventions on hemodynamic, biochemical, and pathological changes were assessed. Subcellular localization of PKC isoforms by Western blot analysis and immunocytochemistry demonstrated that PKC-alpha and PKC-delta were translocated to the sarcolemma and that PKC-delta was translocated to the mitochondria after diazoxide treatment. In hearts treated with diazoxide (80 micromol/L), a significant improvement in cardiac function and an attenuation of cell injury were observed. In PKC-downregulated hearts, protection was abolished because mitoK(ATP) channels could not be activated by diazoxide.

CONCLUSIONS:

These data suggest that PKC activation is required for the opening of mitoK(ATP) channels during protection against ischemia and that this effect is linked to isoform-specific translocation of PKC-delta to the mitochondria.

• Authors: Y Wang, E Takashi, M Xu, A Ayub, M Ashraf
• Journal: Circulation (Circulation) Vol. 104 Issue 1 Pg. 85-90 (Jul 03 2001) ISSN: 1524-4539 [Electronic] United States
• PMID: 11435343 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Isoenzymes
• Potassium Channel Blockers
• Potassium Channels
• Adenosine Triphosphate
• L-Lactate Dehydrogenase
• Protein Kinase C
• Tetradecanoylphorbol Acetate
• Diazoxide

Topics

• Adenosine Triphosphate (metabolism)
• Animals
• Diazoxide (pharmacology)
• Dose-Response Relationship, Drug
• Down-Regulation (drug effects)
• Immunohistochemistry
• In Vitro Techniques
• Ischemic Preconditioning, Myocardial (methods)
• Isoenzymes (metabolism)
• L-Lactate Dehydrogenase (metabolism)
• Male
• Mitochondria (metabolism)
• Myocardial Ischemia (metabolism)
• Myocardium (cytology, enzymology)
• Potassium Channel Blockers
• Potassium Channels (metabolism)
• Protein Kinase C (metabolism)
• Protein Transport (drug effects, physiology)
• Rats
• Rats, Sprague-Dawley
• Reperfusion Injury (metabolism)
• Sarcolemma (metabolism)
• Subcellular Fractions (enzymology)
• Tetradecanoylphorbol Acetate (pharmacology)
• Ventricular Function, Left (drug effects)