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Am J Physiol Heart Circ Physiol 284: H141-H150, 2003. First published October 10, 2002; doi:10.1152/ajpheart.00132.2002
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Vol. 284, Issue 1, H141-H150, January 2003

Reperfusion, not simulated ischemia, initiates intrinsic apoptosis injury in chick cardiomyocytes

Terry L. Vanden Hoek*, Yimin Qin*, Kim Wojcik, Chang-Qing Li, Zuo-Hui Shao, Travis Anderson, Lance B. Becker, and Kimm J. Hamann

Sections of Emergency Medicine and Pulmonary/Critical Care, Department of Medicine and Emergency Resuscitation Center, University of Chicago, Chicago, Illinois 60637

Although ischemia-reperfusion (I/R) can initiate apoptosis, the timing and contribution of the mitochondrial/cytochrome c apoptosis death pathway to I/R injury is unclear. We studied the timing of cytochrome c release during I/R and whether subsequent caspase activation contributes to reperfusion injury in confluent chick cardiomyocytes. One-hour simulated ischemia followed by 3-h reperfusion resulted in significant cell death, with most cell death evident during the reperfusion phase and demonstrating mitochondrial cytochrome c release within 5 min after reperfusion. By contrast, cells exposed to prolonged ischemia for 4 h had only marginally increased cell death and no detectable cytochrome c release into the cytosol. Caspase activation could not be detected after ischemia only, but it significantly increased after reperfusion. Caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, Ac-Asp-Gln-Thr-Asp-H, or benzyloxycarbonyl-Leu-Glu (Ome)-His-Asp-(Ome)-fluoromethyl ketone given only at reperfusion significantly attenuated cell death and resulted in return of contraction. Antixoxidants decreased cytochrome c release, nuclear condensation, and cell death. These results suggest that reperfusion oxidants initiate cytochrome c release within minutes, and apoptosis within hours, significant enough to increase cell death and contractile dysfunction.

oxidants; ischemia-reperfusion; reperfusion injury; caspase inhibition

* T. L. Vanden Hoek and Y. Qin contributed equally to this work.




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