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Oxidised proteins and superoxide anion production in the diaphragm of severe COPD patients

¹ Muscle Research and Respiratory System Unit, Respiratory Medicine and Thoracic Surgery Depts, IMIM-Hospital del Mar, Barcelona, ² School of Medicine, Universitat Autònoma de Barcelona, ⁵ Health and Experimental Science Department (CEXS), Universitat Pompeu Fabra, Barcelona, and ⁴ Centro de Investigación en Red de Enfermedades Respiratorias (CIBERES), Spain. ³ Critical Care and Respiratory Divisions, Royal Victoria Hospital and Meakins-Christie Laboratories, McGill University, Montreal, QC, Canada.

CORRESPONDENCE: E. Barreiro, Muscle and Respiratory System Research Unit, IMIM-Hospital del Mar, PRBB, Dr. Aiguader 88, E-08003 Barcelona, Spain. Fax: 34 933160410. E-mail: ebarreiro{at}imim.es

Keywords: Chronic obstructive pulmonary disease, muscle compartments, oxidised proteins, respiratory muscles, superoxide anion

Received: May 10, 2008
Accepted January 17, 2009

In the diaphragms of chronic obstructive pulmonary disease (COPD) patients, the nature of oxidatively modified proteins and superoxide anion production were explored.

Diaphragm specimens were obtained through thoracotomy because of localised lung lesions in COPD patients (16 severe and eight moderate) and 10 control subjects. Lung and respiratory muscle functions were evaluated. Oxidised proteins were identified using immunoblotting and mass spectrometry. Protein and activity levels of the identified proteins were determined using immunoblotting and activity assays. Lucigenin-derived chemiluminescence signals in a luminometer were used to determine superoxide anion levels in muscle compartments (mitochondria, membrane and cytosol) using selective inhibitors.

In severe COPD patients compared with controls, respiratory muscle function was impaired; creatine kinase, carbonic anhydrase III, actin and myosin were oxidised; myosin carbonylation levels were increased five-fold; creatine kinase content and activity and myosin protein were reduced; superoxide anion levels were increased in both mitochondria and membrane compartments; and the percentage of superoxide anion inhibition achieved by rotenone was significantly greater.

In severe COPD patients, oxidation of diaphragm proteins involved in energy production and contractile performance is likely to partially contribute to the documented respiratory muscle dysfunction. Furthermore, generation of the superoxide anion was increased in the diaphragms of these patients.

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