Src tyrosine kinase is the trigger but not the mediator of ischemic preconditioning.
The signal cascade that triggers and mediates ischemic preconditioning (IPC) remains unclear. The present study investigated the role of the Src family of tyrosine kinases in IPC. Isolated and buffer-perfused rat hearts underwent IPC with three cycles of 5-min ischemia and 5-min reperfusion, followed by 30-min ischemia and 120-min reperfusion. The Src tyrosine kinase family-selective inhibitor PP1 was administered between 45 and 30 min before ischemia (early PP1 treatment) or for 15 min before IPC [early PP1-preconditioning (PC) treatment]. PP1 was also administered for 5 min before the sustained ischemia (late PP1 treatment) or after IPC (late PP1-PC treatment). Src kinase was activated after 30 min of ischemia in both the membrane and cytosolic fractions. Src kinase was also activated by IPC but was attenuated after the sustained ischemia. Early and late PP1 treatment inhibited Src activation after the sustained ischemia and reduced infarct size. Early PP1-PC inhibited Src activation after IPC but not after the sustained ischemia and blocked cardioprotection afforded by IPC. Late PP1-PC treatment abrogated IPC-induced activation of Src and protein kinase C (PKC)-epsilon in the membrane but not in the cytosolic fraction. This treatment modality abrogated Src activation after the sustained ischemia and failed to block cardioprotection afforded by IPC. These results suggest that Src kinase activation mediates ischemic injury but triggers IPC in the position either upstream of or parallel to membrane-associated PKC-epsilon.
Hattori
R
R
Cardiovascular Division, Department of Surgery, University of Connecticut School of Medicine, Farmington, Connecticut 06030, USA.
Otani
H
H
Uchiyama
T
T
Imamura
H
H
Cui
J
J
Maulik
N
N
Cordis
G A
GA
Zhu
L
L
Das
D K
DK
D000818:Animals; D015153:Blotting, Western; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D019157:Ischemic Preconditioning, Myocardial / Q000379:methods; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D017202:Myocardial Ischemia / Q000201:enzymology / Q000473:pathology; D015425:Myocardial Reperfusion / Q000379:methods; D009206:Myocardium / Q000201:enzymology* / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D051744:Protein Kinase C-epsilon; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function / Q000187:drug effects; D015398:Signal Transduction / Q000502:physiology*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
C108123:4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D011720:Pyrazoles; D011743:Pyrimidines; C497257:Prkce protein, rat; D019061:src-Family Kinases; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon