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Ischemia/reperfusion‐induced necrosis and apoptosis in the cells isolated from rat skeletal muscle
Author(s) -
Wang Wei Z.,
Fang XinHua,
Stephenson Linda L.,
Khiabani Kayvan T.,
Zamboni William A.
Publication year - 2008
Publication title -
journal of orthopaedic research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.041
H-Index - 155
eISSN - 1554-527X
pISSN - 0736-0266
DOI - 10.1002/jor.20493
Subject(s) - necrosis , propidium iodide , apoptosis , skeletal muscle , ischemia , annexin , programmed cell death , pi , myocyte , endocrinology , biology , medicine , andrology , pathology , biochemistry
Abstract 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. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:351–356, 2008