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1Departments of Exercise and Sport Sciences and Physiology, Center for Exercise Science, University of Florida, Gainesville, Florida 32611; and 2Faculty of Medicine, Sports Medicine Department, Ankara University, 06100 Tandogan, Ankara, Turkey
Submitted 10 September 2002 ; accepted in final form 16 May 2003
Prolonged mechanical ventilation (MV) results in oxidative damage in the
diaphragm; however, it is unclear whether this MV-induced oxidative injury
occurs rapidly or develops slowly over time. Furthermore, it is unknown
whether both soluble (cytosolic) and insoluble (myofibrillar) proteins are
equally susceptible to oxidation during MV. These experiments tested two
hypotheses: 1) MV-induced oxidative injury in the diaphragm occurs
within the first 6 h after the initiation of MV; and 2) MV is
associated with oxidative modification of both soluble and insoluble proteins.
Adult Sprague-Dawley rats were randomly divided into one of seven experimental
groups: 1) control (n = 8); 2) 3-h MV (n =
8); 3) 6-h MV (n = 6); 4) 18-h MV (n = 8);
5) 3-h anesthesia-spontaneous breathing (n = 8); 6)
6-h anesthesia-spontaneous breathing (n = 6); and 7) 18-h
anesthesia-spontaneous breathing (n = 8). Markers of oxidative injury
in the diaphragm included the measurement of reactive (protein) carbonyl
derivatives (RCD) and total lipid hydroperoxides. Three hours of MV did not
result in oxidative injury in the diaphragm. In contrast, both 6 and 18 h of
MV promoted oxidative injury in the diaphragm, as indicated by increases in
both protein RCD and lipid hydroperoxides. Electrophoretic separation of
soluble and insoluble proteins indicated that the MV-induced accumulation of
RCD was limited to insoluble proteins with molecular masses of 
200, 120,
80, and 40 kDa. We conclude that MV results in a rapid onset of oxidative
injury in the diaphragm and that insoluble proteins are primary targets of
MV-induced protein oxidation.
respiratory muscles; muscle atrophy; free radicals; protein oxidation; weaning from mechanical ventilation; muscle injury
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