title: A comprehensive analysis of oxidative stress in the ozone-induced lung inflammation mouse model.
abstract: Ozone is an oxidizing environmental pollutant that contributes significantly to respiratory health. Exposure to increased levels of ozone has been associated with worsening of symptoms of patients with asthma and COPD (chronic obstructive pulmonary disease). In the present study, we investigated the acute and chronic effects of ozone exposure-induced oxidative stress-related inflammation mechanics in mouse lung. In particular, we investigated the oxidative stress-induced effects on HDAC2 (histone deacetylase 2) modification and activation of the Nrf2 (nuclear factor erythroid-related factor 2) and HIF-1α (hypoxia-inducible factor-1α) signalling pathways. Male C57BL/6 mice were exposed to ozone (3 p.p.m.) for 3 h a day, twice a week for a period of 1, 3 or 6 weeks. Control mice were exposed to normal air. After the last exposure, mice were killed for BAL (bronchoalveolar lavage) fluid and lung tissue collection. BAL total cell counts were elevated at all of the time points studied. This was associated with increased levels of chemokines and cytokines in all ozone-exposed groups, indicating the presence of a persistent inflammatory environment in the lung. Increased inflammation and Lm (mean linear intercept) scores were observed in chronic exposed mice, indicating emphysematous changes were present in lungs of chronic exposed mice. The antioxidative stress response was active (indicated by increased Nrf2 activity and protein) after 1 week of ozone exposure, but this ability was lost after 3 and 6 weeks of ozone exposure. The transcription factor HIF-1α was elevated in 3- and 6-week ozone-exposed mice and this was associated with increased gene expression levels of several HIF-1α target genes including Hdac2 (histone deacetylase 2), Vegf (vascular endothelial growth factor), Keap1 (kelch-like ECH-associated protein 1) and Mif (macrophage migration inhibitory factor). HDAC2 protein was found to be phosphorylated and carbonylated in nuclear and cytoplasm fractions, respectively, and was associated with a decrease in DNA-binding activity and protein expression of HDAC2. Decreased HDAC2 activity, most likely a direct result of protein modification, in combination with the loss of the antioxidative stress response and activation of the HIF-1α pathway, contribute to the inflammatory response and emphysema observed in ozone-exposed mice.
authors.affiliation: *Airways Disease Section, National Heart & Lung Institute, Imperial College London, London SW3 6LY, U.K.
authors.forename: Coen H
authors.initials: CH
authors.lastname: Wiegman
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Clarke
authors.forename: Elen
authors.initials: E
authors.lastname: Jazrawi
authors.forename: Paul
authors.initials: P
authors.lastname: Kirkham
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Barnes
authors.forename: Ian M
authors.initials: IM
authors.lastname: Adcock
authors.forename: Kian F
authors.initials: KF
authors.lastname: Chung
mesh_terms: D000368:Aged; D000393:Air Pollutants / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002452:Cell Count; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D004195:Disease Models, Animal; D005786:Gene Expression Regulation / Q000187:drug effects; D056464:Histone Deacetylase 2 / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D010083:Oxidants, Photochemical / Q000008:administration & dosage / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects*; D010126:Ozone / Q000008:administration & dosage / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011014:Pneumonia / Q000139:chemically induced* / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D011656:Pulmonary Emphysema / Q000139:chemically induced; D012333:RNA, Messenger / Q000235:genetics; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D000393:Air Pollutants; D000975:Antioxidants; D016207:Cytokines; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D018836:Inflammation Mediators; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D010083:Oxidants, Photochemical; D012333:RNA, Messenger; D010126:Ozone; D013482:Superoxide Dismutase; C533424:Hdac2 protein, mouse; D056464:Histone Deacetylase 2