title: Amifostine reduces lung vascular permeability via suppression of inflammatory signalling.
abstract: Despite an encouraging outcome of antioxidant therapy in animal models of acute lung injury, effective antioxidant agents for clinical application remain to be developed. The present study investigated the effect of pre-treatment with amifostine, a thiol antioxidant compound, on lung endothelial barrier dysfunction induced by Gram-negative bacteria wall-lipopolysaccharide (LPS). Endothelial permeability was monitored by changes in transendothelial electrical resistance. Cytoskeletal remodelling and reactive oxygen species (ROS) production was examined by immunofluorescence. Cell signalling was assessed by Western blot. Measurements of Evans blue extravasation, cell count and protein content in bronchoalveolar lavage fluid were used as in vivo parameters of lung vascular permeability. Hydrogen peroxide, LPS and interleukin-6 caused cytoskeletal reorganisation and increased permeability in the pulmonary endothelial cells, reflecting endothelial barrier dysfunction. These disruptive effects were inhibited by pre-treatment with amifostine and linked to the amifostine-mediated abrogation of ROS production and redox-sensitive signalling cascades, including p38, extracellular signal regulated kinase 1/2, mitogen-activated protein kinases and the nuclear factor-kappaB pathway. In vivo, concurrent amifostine administration inhibited LPS-induced oxidative stress and p38 mitogen-activated protein kinase activation, which was associated with reduced vascular leak and neutrophil recruitment to the lungs. The present study demonstrates, for the first time, protective effects of amifostine against lipopolysaccharide-induced lung vascular leak in vitro and in animal models of lipopolysaccharide-induced acute lung injury.
authors.affiliation: Section of Pulmonary and Critical Care Medicine, Department of Medicine, University of Chicago, 929 E. 57th Street, GCIS Bldg, Chicago, IL 60637, USA.
authors.forename: P
authors.initials: P
authors.lastname: Fu
authors.forename: A A
authors.initials: AA
authors.lastname: Birukova
authors.forename: J
authors.initials: J
authors.lastname: Xing
authors.forename: S
authors.initials: S
authors.lastname: Sammani
authors.forename: J S
authors.initials: JS
authors.lastname: Murley
authors.forename: J G N
authors.initials: JG
authors.lastname: Garcia
authors.forename: D J
authors.initials: DJ
authors.lastname: Grdina
authors.forename: K G
authors.initials: KG
authors.lastname: Birukov
mesh_terms: D004999:Amifostine / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid; D002199:Capillary Permeability / Q000187:drug effects*; D003599:Cytoskeleton / Q000378:metabolism; D007249:Inflammation; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011837:Radiation-Protective Agents / Q000494:pharmacology*; D017382:Reactive Oxygen Species; D015398:Signal Transduction
chemical_list: D000975:Antioxidants; D015850:Interleukin-6; D008070:Lipopolysaccharides; D011837:Radiation-Protective Agents; D017382:Reactive Oxygen Species; D004999:Amifostine