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1 Departments of Exercise and Sport Sciences and Physiology, University of Florida, Gainesville, FL, USA; Faculty of Medicine, Sports Medicine Department, Ankara University, Ankara, Turkey
2 Departments of Exercise and Sport Sciences and Physiology, University of Florida, Gainesville, FL, USA
* To whom correspondence should be addressed. E-mail: spowers{at}hhp.ufl.edu.
Prolonged mechanical ventilation (MV) results in oxidative damage in the diaphragm; however, it is unclear if this MV-induced oxidative injury occurs rapidly or develops slowly over time. Further, it is unknown if 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 hours 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 hours MV (n=8); 3) 6 hours MV (n=6); 4) 18 hours MV (n=8); 5) 3 hours anesthesia-spontaneous breathing (n=8); 6) 6 hours anesthesia-spontaneous breathing (n=6); and 7) 18 hours 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 hours 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 weights around 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.
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