Fas-induced pulmonary apoptosis and inflammation during indirect acute lung injury

Am J Respir Crit Care Med. 2007 Sep 15;176(6):591-601. doi: 10.1164/rccm.200611-1743OC. Epub 2007 Jun 28.

Abstract

Rationale: Indirect acute lung injury (ALI) is associated with high morbidity and mortality. No specific therapies have been developed, because the underlying pathophysiological processes remain elusive.

Objectives: To investigate the contribution of Fas-induced apoptotic and nonapoptotic/inflammatory signaling to the pathology of indirect ALI.

Methods: A mouse model of indirect ALI, induced by successive exposure to hemorrhagic shock and cecal ligation and puncture, was used. Quantification of active caspase-3 and the short splice variant of FLICE-inhibitory protein, (FLIP)short, was performed by Western blotting and immunohistochemistry, and cytokines/chemokines were assessed by cytometric bead array or ELISA. M30 immunostaining was done to evaluate epithelial cell apoptosis. Lung injury was assessed on the basis of myeloperoxidase activity, bronchoalveolar lavage protein, and lung histology.

Measurements and main results: Twelve hours after insult, lung monocyte chemoattractant protein-1, keratinocyte-derived chemokine, macrophage inflammatory protein-2, IL-6, tumor necrosis factor-alpha, and caspase-3 were increased and FLIP(short) was decreased. Fas- and Fas ligand-deficient mice showed marked protection from lung inflammation and apoptosis and decreased ALI. This was associated with a 10-day survival benefit. Similarly, 4 hours after pulmonary instillation of Fas-activating antibody in vivo, lung chemokines were markedly elevated in background mice and, interestingly, to a similar degree in macrophage-deficient animals. Fas activation on lung epithelial cells in vitro led to chemokine production that was dependent on extracellular signal-regulated kinase.

Conclusions: Activation of apoptotic and nonapoptotic/inflammatory Fas signaling is an early important pathophysiological event in the development of indirect ALI after hemorrhagic shock and sepsis, in which lung epithelial cells appear to play a central role.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Antibodies, Monoclonal / toxicity
  • Apoptosis*
  • Blotting, Western
  • Bronchoalveolar Lavage Fluid / chemistry
  • CASP8 and FADD-Like Apoptosis Regulating Protein / metabolism
  • Caspase 3 / metabolism
  • Cell Line
  • Cell Proliferation
  • Disease Models, Animal
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Male
  • Mice
  • Mice, Mutant Strains
  • Prognosis
  • Respiratory Distress Syndrome / chemically induced
  • Respiratory Distress Syndrome / metabolism
  • Respiratory Distress Syndrome / pathology*
  • Severity of Illness Index
  • fas Receptor / immunology
  • fas Receptor / toxicity*

Substances

  • Actins
  • Antibodies, Monoclonal
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • Fas protein, mouse
  • fas Receptor
  • Caspase 3