Elsevier

Nitric Oxide

Volume 9, Issue 1, August 2003, Pages 33-43
Nitric Oxide

Bronchoalveolar lavage fluids of ventilated patients with acute lung injury activate NF-κB in alveolar epithelial cell line: role of reactive oxygen/nitrogen species and cytokines

Communicated by Ulrich Forstermann
https://doi.org/10.1016/j.niox.2003.07.001Get rights and content

Abstract

In human alveolar epithelial cell line, we investigated the binding activity of NF-κB 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-1β, IL-8, and nitrated proteins (NTP), the activity of myeloperoxidase, and the capacity to activate the NF-κB in alveolar A549 cells by electrophoretic mobility shift and supershift assays. The neutrophil counts and mean IL-1β, 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-1β, IL-8, myeloperoxidase, NTP, and endotoxin in all groups (P<0.0001). NF-κB 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-1β (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-κB-binding activity. These mediators and NF-κB 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-κB activation.

Section snippets

Patients

Sixty-seven patients from the general ICU of the University Hospital Center in Liège with clinical suspicion of nosocomial pneumonia were included in the study. All patients were intubated and mechanically ventilated for >72 h for multiple reasons, such as multiple or cerebral trauma, emergency neurosurgery, complicated abdominal surgery, burns, miscellaneous infections, hematologic malignancies, or post-cardiovascular surgery complications (Table 1). As a routine practice, BAL was performed

General data

Table 1 summarizes the population characteristics and clinical parameters of the 67 patients included in the study. Bronchopneumonia was confirmed by quantitative culture in 30 patients. Among the 37 patients without bronchopneumonia, 20 were in ARDS of varying etiology. Therefore, patients were divided into four groups: patients with bronchopneumonia (n=16, BALs=16, Group 1), patients in ARDS without pneumonia (n=20, BALs=22, Group 2), patients in ARDS with pneumonia (n=14, BALs=17, Group 3),

Discussion

We previously demonstrated that nitrated proteins (NTP) were present in BAL fluids of ventilated ICU patients, correlated to the oxidant activity of neutrophils, and that these BAL fluids were toxic for alveolar like cells in culture [9]. In this study, we demonstrate that BAL fluid specimens (as a reflect of alveolar milieu) of mechanically ventilated patients are able to stimulate the NF-κB activity in pneumocytes in culture (A549 cells), especially BAL fluids of patients with

Acknowledgements

The authors gratefully acknowledge the skilful technical assistance of the technical staff of the Center of Oxygen, Research and Development. This work was supported by Grant 3.4542.00 from the National Fund for Scientific Research (Belgium) and the Fund of Investments for Scientific Research of University Hospital of Liège (Belgium).

References (45)

  • P.M Suter et al.

    High bronchoalveolar levels of tumor necrosis factor and its inhibitors, interleukin-1, interferon, and elastase, in patients with adult respiratory distress syndrome after trauma, shock, or sepsis

    Am. Rev. Respir. Dis.

    (1992)
  • C.G Cochrane

    Cellular injury by oxidants

    Am. J. Med.

    (1991)
  • J.S Beckman et al.

    Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide

    Proc. Natl Acad. Sci. USA

    (1990)
  • N.W Kooy et al.

    Evidence for in vivo peroxynitrite production in human acute lung injury

    Am. J. Respir. Crit. Care Med.

    (1995)
  • M Mathy-Hartert et al.

    Nitrated proteins in bronchoalveolar lavage fluid of patients at risk of ventilator-associated bronchopneumonia

    Eur. Respir. J.

    (2000)
  • S Ghosh et al.

    NF-κB and Rel proteins: evolutionarily conserved mediators of immune responses

    Annu. Rev. Immunol.

    (1998)
  • F Arenzana-Seisdedos et al.

    Nuclear localization of I kappa B alpha promotes active transport of NF-kappa B from the nucleus to the cytoplasm

    J. Cell. Sci.

    (1997)
  • M Meyer et al.

    H2O2 and antioxidants have opposite effects on activation of NF-κB and AP-1 in intact cells: AP-1 as secondary antioxidant-responsive factor

    EMBO J.

    (1993)
  • A.A Beg et al.

    Tumor necrosis factor and interleukin-1 lead to phosphorylation and loss of IκBα: a mechanism for NF-κB activation

    Mol. Cell. Biol.

    (1993)
  • P.J Barnes et al.

    Nuclear factor-κB: a pivotal transcription factor in chronic inflammatory diseases

    N. Engl. J. Med.

    (1997)
  • M.D Schwartz et al.

    Nuclear factor-κB is activated in alveolar macrophages from patients with acute respiratory distress syndrome

    Crit. Care Med.

    (1996)
  • P Moine et al.

    NF-κB regulatory mechanisms in alveolar macrophages from patients with acute respiratory distress syndrome

    Shock

    (2000)

    • Cited by (0)

    View full text
    Copyright © 2003 Elsevier Inc. All rights reserved.