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1 Cardiovascular Division, Department of Surgery, University of Connecticut School of Medicine, Farmington, Connecticut 06030; and 2 Department of Thoracic and Cardiovascular Surgery, Kansai Medical University, Moriguchi City 570-8507, Japan
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)-
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-.
protein kinase C-; PP1
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