title: The effect of oxidative stress on macrophages and lung epithelial cells: the role of phosphodiesterases 1 and 4.
abstract: Reactive oxygen species (ROS) have been implicated in various pulmonary diseases by causing direct injury to lung epithelial cells. Signalling activity of cells through transcription factors such as nuclear factor kappa B (NF-kappaB) and AP-1 have been shown to be regulated by ROS, and the release of pro-inflammatory cytokines demonstrated in the study of inflammatory disease. In this study, we examined the effect of the oxidant tert-butylhydroperoxide (tBHP) on mouse J774 macrophages and its ability to cause the release of the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-alpha). The role of calcium as a signalling molecule was studied using various calcium antagonists. The role of the signalling molecule cAMP was also investigated using phosphodiesterase inhibitors PDE1 and PDE4 families. Oxidative stress was investigated in lung epithelial (A549) cells with and without calcium antagonists and PDE inhibitors with regard to their ability to modulate release of the neutrophil chemoattractant interleukin 8 (IL-8). The oxidant tBHP significantly increased the cytosolic calcium concentration in J774 macrophages, which was prevented by the PDE1 inhibitor. The production of TNF-alpha protein by J774 macrophages was mediated by a pathway involving calcium as addition of calcium antagonists inhibited the tBHP stimulated increase in the cytokine. Inhibitors of both PDE1 and PDE4 completely prevented the tBHP stimulated TNF-alpha release suggesting that the cAMP pathway may be important in the oxidant induced signalling pathway leading to gene expression of pro-inflammatory cytokines. In the presence of oxidant alone, A549 epithelial cells released significant amounts of IL-8, which was inhibited by both calcium antagonist treatment and PDE inhibition treatment. These data suggest that ROS-mediated lung inflammation could be mediated at least in part by calcium and elevated PDE activity associated with decreased cAMP in both macrophages and epithelial cells. Inhibition of these pathways may provide a route for treatment of inflammatory lung diseases.
authors.affiliation: School of Life Sciences, Napier University, 10 Colinton Road, Edinburgh EH10 5DT, UK. da.brown@napier.ac.uk
authors.forename: D M
authors.initials: DM
authors.lastname: Brown
authors.forename: L
authors.initials: L
authors.lastname: Hutchison
authors.forename: K
authors.initials: K
authors.lastname: Donaldson
authors.forename: S J
authors.initials: SJ
authors.lastname: MacKenzie
authors.forename: C A J
authors.initials: CA
authors.lastname: Dick
authors.forename: V
authors.initials: V
authors.lastname: Stone
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors; D012368:4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008168:Lung / Q000166:cytology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D016877:Oxidants / Q000494:pharmacology*; D018384:Oxidative Stress; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D013449:Sulfonamides / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology; D020122:tert-Butylhydroperoxide / Q000494:pharmacology*
chemical_list: D002121:Calcium Channel Blockers; D002614:Chelating Agents; D016209:Interleukin-8; D016877:Oxidants; D010726:Phosphodiesterase Inhibitors; D013449:Sulfonamides; D014409:Tumor Necrosis Factor-alpha; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D012368:4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; D004533:Egtazic Acid; C017967:W 7; D020122:tert-Butylhydroperoxide; D014700:Verapamil; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D002118:Calcium