Carbonylated proteins in bronchoalveolar lavage of patients with sarcoidosis, pulmonary fibrosis associated with systemic sclerosis and idiopathic pulmonary fibrosis.
Oxygen-derived free radicals produced by phagocytes have been postulated to contribute to lung tissue damage occurring during diffuse lung diseases (DLD). The two-dimensional electrophoretic (2-DE) analysis of bronchoalveolar lavage (BAL) protein composition revealed different protein profiles in sarcoidosis (S), idiopathic pulmonary fibrosis (IPF) and systemic sclerosis (SSc) with a significant increase of low molecular weight proteins in IPF. Some of these proteins are involved in antioxidant processes. The aims of this report were to analyse the oxidative stress occurring in patients with DLD through determination of BAL protein carbonyl content and to identify target proteins of oxidation by a proteomic approach (2-DE combined with immunoblotting with specific antibodies for carbonyl groups). Carbonylated proteins detected by enzyme-linked immunosorbent assay (ELISA) were increased in BAL of patients with S, IPF and SSc compared to healthy controls with a significant difference for S and IPF. The proteomic approach to the analysis of BAL revealed that protein carbonylation was a process involving specific carbonylation-sensitive proteins and that in IPF a greater number of proteins target of oxidation were present. In conclusion, to our knowledge, this is the first report providing a database of proteins target of oxidation in BAL of patients with sarcoidosis, idiopathic pulmonary fibrosis and systemic sclerosis.
Rottoli
Paola
P
Respiratory Diseases Section, Department of Clinical Medicine and Immunological Sciences, University of Siena, Siena, Italy. rottoli@unisi.it
Magi
Barbara
B
Cianti
Riccardo
R
Bargagli
Elena
E
Vagaggini
Cecilia
C
Nikiforakis
Nikolaos
N
Pallini
Vitaliano
V
Bini
Luca
L
D015153:Blotting, Western; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011658:Pulmonary Fibrosis / Q000209:etiology / Q000378:metabolism*; D012016:Reference Values; D012507:Sarcoidosis / Q000378:metabolism*; D012595:Scleroderma, Systemic / Q000150:complications*
D011506:Proteins