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Neutrophil mediated muscle injury of the diaphragm Samrai, Baljit


Excessive loading, as seen in chronic respiratory diseases and experimentally produced by tracheal banding (TB) may result in injury to the myofibres of the diaphragm and hypercapnic ventilatory failure. Neutrophils (PMNs) are primary inflammatory cells that migrate to an area of injury in order to phagocytose cellular debris and promote the inflammatory response. During this process, neutrophils may also amplify muscle injury. We hypothesized that neutrophil depletion would reduce diaphragm muscle injury in an animal model where diaphragm injury was induced by tracheal banding. Adult male Sprague-Dawley rats were randomly assigned to one of four groups: 1. control (C), 2. tracheal banded (TB), 3. control plus neutrophil depleted (C+ND), 4. tracheal banded plus neutrophil depleted (TB+ND). In the C+ND and TB+ND groups, neutrophils were depleted by administering anti-rat polymorphonuclear leukocyte antisera, beginning one day pre-surgery. After anaesthesia, the carotid artery was chronically cannulated, esophageal pressures (Pes) were measured and a cuff was tightened around the isolated trachea, until the Pes during tidal breathing (Pes[sub TV]) was 20% of the maximal Pes (Pes[sub Max]) during tracheal occlusion in the TB and TB+ND groups. Arterial blood gas (ABG) samples were taken daily from the awake animal. Three days later, following anaesthesia, in vivo measures of Pes were repeated and then the diaphragm and trachea were excised for histological and immunohistochemical analysis. The following parameters were investigated: (1) the presence of neutrophils in the diaphragm measured by neutrophil specific myeloperoxidase (MPO) activity and point counting H&E diaphragm cross-sections; (2) muscle injury as quantified by point counting H&E stained diaphragm cross-sections; (3) the presence of EDI positive macrophages in the diaphragm by using immunohistochemistry; and (4) ABGs measure daily post-surgery. The TB groups were hypercapnic (PaC0₂ > 45mmHg; p

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