Bronchoalveolar lavage fluids of ventilated patients with acute lung injury activate NF-kappaB in alveolar epithelial cell line: role of reactive oxygen/nitrogen species and cytokines.
In human alveolar epithelial cell line, we investigated the binding activity of NF-kappaB induced by the bronchoalveolar lavage fluids (BALs) from ventilated patients with acute lung injury (ALI), in correlation with the concentrations of inflammatory cytokines, RNOS, and the severity of the ALI. In BALs obtained in 67 patients (16 bronchopneumonia, 14 infected ARDS, 20 ARDS, and 17 ALI patients without bronchopneumonia and no ARDS), we measured endotoxin, IL-1beta, IL-8, and nitrated proteins (NTP), the activity of myeloperoxidase, and the capacity to activate the NF-kappaB in alveolar A549 cells by electrophoretic mobility shift and supershift assays. The neutrophil counts and mean IL-1beta, IL-8, myeloperoxidase, and NTP values were increased in bronchopneumonia and infected ARDS groups compared to ARDS and ALI without bronchopneumonia and no ARDS groups (P<0.001). The number of neutrophils was correlated to those of IL-1beta, IL-8, myeloperoxidase, NTP, and endotoxin in all groups (P<0.0001). NF-kappaB activity was induced in alveolar like cells by BALs in all groups, was higher in bronchopneumonia and infected ARDS groups (P<0.02), and was correlated to IL-1beta (P=0.0002), IL-8 (P=0.02), NTP (P=0.014), myeloperoxidase (P=0.016), and neutrophil counts (P=0.003). BALs of bronchopneumonia and infected ARDS patients had increased inflammatory mediators (compared to ARDS and ALI without bronchopneumonia and no ARDS patients) that correlated to neutrophil counts and to the NF-kappaB-binding activity. These mediators and NF-kappaB activation may induce an amplification of inflammatory phenomena. By in vitro studies, we confirmed that NO-derived species (10(-6) to 10(-5)M peroxynitrite and 10(-5)M nitrites) and myeloperoxidase (at concentration equivalent to that found in BALs) can participate in the NF-kappaB activation.
Nys
M
M
Department of Anesthesia and Intensive Care Medicine, University of Liège, B4000 Sart Tilman, Liège, Belgium. Monique.Nys@chu.ulg.ac.be
Deby-Dupont
G
G
Habraken
Y
Y
Legrand-Poels
S
S
Kohnen
S
S
Ledoux
D
D
Canivet
J-L
JL
Damas
P
P
Lamy
M
M
D000208:Acute Disease; D000328:Adult; D000368:Aged; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D001996:Bronchopneumonia / Q000378:metabolism; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D004731:Endotoxins / Q000032:analysis / Q000494:pharmacology; D006801:Humans; D007378:Interleukins / Q000032:analysis / Q000494:pharmacology; D008168:Lung / Q000737:chemistry / Q000378:metabolism; D055370:Lung Injury; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism*; D018375:Neutrophil Activation; D009574:Nitro Compounds / Q000032:analysis / Q000494:pharmacology; D009195:Peroxidase / Q000032:analysis / Q000494:pharmacology; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000378:metabolism*; D026361:Reactive Nitrogen Species / Q000378:metabolism* / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012128:Respiratory Distress Syndrome / Q000378:metabolism
D016207:Cytokines; D004731:Endotoxins; D007378:Interleukins; D016328:NF-kappa B; D009574:Nitro Compounds; D011506:Proteins; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D009195:Peroxidase