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.
Wang
Wei Z
WZ
Department of Surgery, University of Nevada School of Medicine, 2040 W. Charleston Blvd., Suite 301, Las Vegas, Nevada 89102, USA. wzwang2000@yahoo.com
Fang
Xin-Hua
XH
Stephenson
Linda L
LL
Khiabani
Kayvan T
KT
Zamboni
William A
WA
D000818:Animals; D017304:Annexin A5; D017209:Apoptosis; D002469:Cell Separation; D004396:Coloring Agents; D004338:Drug Combinations; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D008297:Male; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000473:pathology*; D009336:Necrosis; D011419:Propidium; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000473:pathology* / Q000503:physiopathology*; D013194:Staining and Labeling
D017304:Annexin A5; D004396:Coloring Agents; D004338:Drug Combinations; D005456:Fluorescent Dyes; D011419:Propidium; D016650:Fluorescein-5-isothiocyanate