Ischemia/reperfusion-induced necrosis and apoptosis in the cells isolated from rat skeletal muscle.

Necrosis was considered to be the solo mechanism for ischemia/reperfusion (I/R)-induced cell death. Recent evidence from I/R models of the heart, liver, kidney, and brain indicates that apoptosis is a major contributor to I/R-induced cell death. However, evidence of I/R-induced apoptosis in skeletal muscle is sparse and divided. The purpose for the present study was to investigate I/R-induced necrosis and apoptosis in the cells isolated from rat skeletal muscle. A rat gracilis muscle model was used. After surgical preparation, clamps were applied on the vascular pedicle to create 4 h of ischemia and released for 24 h of reperfusion (I/R, n = 10). Clamping was omitted in sham I/R rats (sham I/R, n = 10). The muscle samples were harvested after 24 h of reperfusion for the process of cell isolation. Cells were stained by Propidium Iodide (PI) or Annexin V-FITC or both. Twenty thousand cells from each muscle sample were scanned and analyzed by flow cytometry. The average percentage of live cells was 45 +/- 2% in the I/R group versus 65 +/- 3% in the sham I/R group (p < 0.01). The average percentage of necrotic cells was 18 +/- 1% in I/R versus 12 +/- 1% in sham I/R (p < 0.01). The average percentage of apoptotic cells was 40 +/- 3% in I/R versus 27 +/- 3% in sham I/R (p < 0.01). Our results clearly demonstrated that I/R not only causes necrosis, but also accelerates apoptosis in the cells isolated from rat skeletal muscle. Copyright 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.