Direct evidence that ischemic preconditioning does not cause protein kinase C translocation in rabbit heart.
OBJECTIVE
Indirect pharmacological evidence suggests that myocardial protection conferred by ischemic preconditioning in rabbit myocardium is mediated through the translocation of protein kinase C (PKC). To test this hypothesis, we performed direct biochemical measurements of subcellular distribution of PKC in rabbit hearts.


METHODS
Two protocols were utilized. In Protocol I the preconditioned group (PC) underwent two 5-min episodes of brief coronary artery occlusion each followed by 5 min of reperfusion, while the control group consisted of time-matched, sham-operated (non-ischemic) animals (SO). Tissue samples were homogenized and cytosolic and particulate fractions were obtained by ultracentrifugation. In Protocol II one group of rabbits received ischemic preconditioning as in Protocol I followed by 10 min of sustained ischemia (PC + IS); a second control group was subjected to 10 min of sustained ischemia (IS); and the third group was time-matched, sham-operated (non-ischemic) animals (SO). Homogenized tissue samples were separated into cytosolic, nuclear and membrane fractions.


RESULTS
In Protocol I, no differences in the subcellular distribution of PKC between the SO and PC groups were observed. In Protocol II, when samples were obtained at 10 min of sustained ischemia, no changes in the subcellular distribution of PKC were observed between SO, IS and PC + IS groups.


CONCLUSIONS
Our results indicate that translocation of protein kinase C is not an important mediator of ischemic preconditioning in the rabbit ischemia model.
Simkhovich
B Z
BZ
Heart Institute, Good Samaritan Hospital, Los Angeles, CA 90017, USA.
Przyklenk
K
K
Hale
S L
SL
Patterson
M
M
Kloner
R A
RA
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D019157:Ischemic Preconditioning, Myocardial; D017202:Myocardial Ischemia / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology; D011493:Protein Kinase C / Q000032:analysis / Q000378:metabolism*; D011817:Rabbits; D011897:Random Allocation
D011493:Protein Kinase C