Aberrant antioxidant activity and excessive deposition of extracellular matrix (ECM) are hallmarks of interstitial lung diseases (ILD). It is known that oxidative stress alters the ECM, but extracellular antioxidant defence mechanisms in ILD are incompletely understood. Here, we extracted abundance and detergent solubility of extracellular antioxidant enzymes from a proteomic dataset of bleomycin-induced lung fibrosis in mice and assessed regulation and distribution of glutathione peroxidase 3 (GPX3) in murine and human lung fibrosis. Superoxide dismutase 3 (Sod3), Gpx3, and Gpx activity were increased in mouse BALF during bleomycin-induced lung fibrosis. In lung tissue homogenates, Gpx3, but not Sod3, was upregulated and detergent solubility profiling indicated that Gpx3 associated with ECM proteins. Immunofluorescence analysis showed that Gpx3 was expressed by bronchial epithelial cells and interstitial fibroblasts and localized to the basement membrane and interstitial ECM in lung tissue. As to human ILD samples, BALF of some patients contained high levels of GPX3, and GPX3 was upregulated in lung homogenates from IPF patients. GPX3 expression in primary human bronchial epithelial cells and lung fibroblasts was downregulated by TNF-α, but more variably regulated by TGF-β1 and menadione. In conclusion, the antioxidant enzyme GPX3 localizes to lung ECM and is variably upregulated in ILD.

Asklepios Fachkliniken München Gauting Munich Germany

Comprehensive Pneumology Center Helmholtz Zentrum München Member of the German Center of Lung Research Munich Germany

Department of Proteomics and Signal Transduction Max Planck Institute of Biochemistry Martinsried Germany

Department of Respiratory Medicine of the 1st Medical School Charles University and Thomayer Hospital Prague Czech republic

Medizinische Klinik und Poliklinik 5 Klinikum der Ludwig Maximilians Universität Member of the German Center for Lung Research Munich Germany

Thoraxchirurgisches Zentrum Klinik für Allgemeine Viszeral Transplantations Gefäß und Thoraxchirurgie Klinikum Großhadern Ludwig Maximilians Universität Munich Germany

Zobrazit více v PubMed

Grunes D. & Beasley M. B. Hypersensitivity pneumonitis: a review and update of histologic findings. J. Clin. Pathol. 66, 888–895, doi: 10.1136/jclinpath-2012-201337 (2013). PubMed DOI

Selman M., Pardo A. & King T. E. Jr. Hypersensitivity pneumonitis: insights in diagnosis and pathobiology. Am. J. Respir. Crit. Care Med. 186, 314–324, doi: 10.1164/rccm.201203-0513CI (2012). PubMed DOI

Katzenstein A. L. Smoking-related interstitial fibrosis (SRIF), pathogenesis and treatment of usual interstitial pneumonia (UIP), and transbronchial biopsy in UIP. Mod. Pathol. 25 Suppl 1, S68–78, doi: 10.1038/modpathol.2011.154 (2012). PubMed DOI

Valeyre D. et al.. Sarcoidosis. Lancet 383, 1155–1167, doi: 10.1016/S0140-6736(13)60680-7 (2014). PubMed DOI

Patterson K. C. & Strek M. E. Pulmonary fibrosis in sarcoidosis. Clinical features and outcomes. Ann. Am. Thorac. Soc. 10, 362–370, doi: 10.1513/AnnalsATS.201303-069FR (2013). PubMed DOI

Kim D. S., Collard H. R. & King T. E. Jr. Classification and natural history of the idiopathic interstitial pneumonias. Proc. Am. Thorac. Soc. 3, 285–292, doi: 10.1513/pats.200601-005TK (2006). PubMed DOI PMC

Raghu G., Weycker D., Edelsberg J., Bradford W. Z. & Oster G. Incidence and prevalence of idiopathic pulmonary fibrosis. Am. J. Respir. Crit. Care Med. 174, 810–816, doi: 10.1164/rccm.200602-163OC (2006). PubMed DOI

Cheresh P., Kim S. J., Tulasiram S. & Kamp D. W. Oxidative stress and pulmonary fibrosis. Biochim. Biophys. Acta 1832, 1028–1040, doi: 10.1016/j.bbadis.2012.11.021 (2013). PubMed DOI PMC

Liu R. M. & Gaston Pravia K. A. Oxidative stress and glutathione in TGF-beta-mediated fibrogenesis. Free Radical Biol. Med. 48, 1–15, doi: 10.1016/j.freeradbiomed.2009.09.026 (2010). PubMed DOI PMC

Maier K., Leuschel L. & Costabel U. Increased levels of oxidized methionine residues in bronchoalveolar lavage fluid proteins from patients with idiopathic pulmonary fibrosis. Am. Rev. Respir. Dis. 143, 271–274, doi: 10.1164/ajrccm/143.2.271 (1991). PubMed DOI

Rottoli P. et al.. Carbonylated proteins in bronchoalveolar lavage of patients with sarcoidosis, pulmonary fibrosis associated with systemic sclerosis and idiopathic pulmonary fibrosis. Proteomics 5, 2612–2618, doi: 10.1002/pmic.200401206 (2005). PubMed DOI

Psathakis K. et al.. Exhaled markers of oxidative stress in idiopathic pulmonary fibrosis. Eur. J. Clin. Invest. 36, 362–367, doi: 10.1111/j.1365-2362.2006.01636.x (2006). PubMed DOI

Bargagli E. et al.. Analysis of carbonylated proteins in bronchoalveolar lavage of patients with diffuse lung diseases. Lung 185, 139–144, doi: 10.1007/s00408-007-9001-6 (2007). PubMed DOI

Rahman I. et al.. Systemic and pulmonary oxidative stress in idiopathic pulmonary fibrosis. Free Radical Biol. Med. 27, 60–68 (1999). PubMed

Cantin A. M., Hubbard R. C. & Crystal R. G. Glutathione deficiency in the epithelial lining fluid of the lower respiratory tract in idiopathic pulmonary fibrosis. Am. Rev. Respir. Dis. 139, 370–372, doi: 10.1164/ajrccm/139.2.370 (1989). PubMed DOI

Meyer A., Buhl R. & Magnussen H. The effect of oral N-acetylcysteine on lung glutathione levels in idiopathic pulmonary fibrosis. Eur. Respir. J. 7, 431–436 (1994). PubMed

Boots A. W. et al.. Antioxidant status associated with inflammation in sarcoidosis: a potential role for antioxidants. Respir. Med. 103, 364–372, doi: 10.1016/j.rmed.2008.10.007 (2009). PubMed DOI

Behr J., Degenkolb B., Beinert T., Krombach F. & Vogelmeier C. Pulmonary glutathione levels in acute episodes of Farmer’s lung. Am. J. Respir. Crit. Care Med. 161, 1968–1971, doi: 10.1164/ajrccm.161.6.9907112 (2000). PubMed DOI

Hecker L. et al.. NADPH oxidase-4 mediates myofibroblast activation and fibrogenic responses to lung injury. Nat. Med. 15, 1077–1081, doi: 10.1038/nm.2005 (2009). PubMed DOI PMC

Carnesecchi S. et al.. A key role for NOX4 in epithelial cell death during development of lung fibrosis. Antioxid. Redox Signal. 15, 607–619, doi: 10.1089/ars.2010.3829 (2011). PubMed DOI PMC

Winterbourn C. C. Reconciling the chemistry and biology of reactive oxygen species. Nat. Chem. Biol. 4, 278–286, doi: 10.1038/nchembio.85 (2008). PubMed DOI

Kagan H. M. & Li W. Lysyl oxidase: properties, specificity, and biological roles inside and outside of the cell. J. Cell. Biochem. 88, 660–672, doi: 10.1002/jcb.10413 (2003). PubMed DOI

Parker M. W. et al.. Fibrotic extracellular matrix activates a profibrotic positive feedback loop. J. Clin. Invest. 124, 1622–1635, doi: 10.1172/JCI71386 (2014). PubMed DOI PMC

Kliment C. R. & Oury T. D. Oxidative stress, extracellular matrix targets, and idiopathic pulmonary fibrosis. Free Radical Biol. Med. 49, 707–717, doi: 10.1016/j.freeradbiomed.2010.04.036 (2010). PubMed DOI

Fattman C. L. et al.. Enhanced bleomycin-induced pulmonary damage in mice lacking extracellular superoxide dismutase. Free Radical Biol. Med. 35, 763–771 (2003). PubMed

Kliment C. R. et al.. Extracellular superoxide dismutase protects against matrix degradation of heparan sulfate in the lung. Antioxid. Redox Signal. 10, 261–268, doi: 10.1089/ars.2007.1906 (2008). PubMed DOI PMC

Kliment C. R. et al.. Oxidative stress alters syndecan-1 distribution in lungs with pulmonary fibrosis. J. Biol. Chem. 284, 3537–3545, doi: 10.1074/jbc.M807001200 (2009). PubMed DOI PMC

Zelko I. N. & Folz R. J. Extracellular superoxide dismutase attenuates release of pulmonary hyaluronan from the extracellular matrix following bleomycin exposure. FEBS Lett. 584, 2947–2952, doi: 10.1016/j.febslet.2010.05.025 (2010). PubMed DOI PMC

Bjermer L., Lundgren R. & Hallgren R. Hyaluronan and type III procollagen peptide concentrations in bronchoalveolar lavage fluid in idiopathic pulmonary fibrosis. Thorax 44, 126–131 (1989). PubMed PMC

Cormier Y., Laviolette M., Cantin A., Tremblay G. M. & Begin R. Fibrogenic activities in bronchoalveolar lavage fluid of farmer’s lung. Chest 104, 1038–1042 (1993). PubMed

Kinnula V. L. Focus on antioxidant enzymes and antioxidant strategies in smoking related airway diseases. Thorax 60, 693–700, doi: 10.1136/thx.2004.037473 (2005). PubMed DOI PMC

Burk R. F., Olson G. E., Winfrey V. P., Hill K. E. & Yin D. Glutathione peroxidase-3 produced by the kidney binds to a population of basement membranes in the gastrointestinal tract and in other tissues. Am. J. Physiol. Gastrointest. Liver Physiol. 301, G32–38, doi: 10.1152/ajpgi.00064.2011 (2011). PubMed DOI PMC

Yamada Y. et al.. Major shifts in the spatio-temporal distribution of lung antioxidant enzymes during influenza pneumonia. PLoS One 7, e31494, doi: 10.1371/journal.pone.0031494 (2012). PubMed DOI PMC

Gao F., Kinnula V. L., Myllarniemi M. & Oury T. D. Extracellular superoxide dismutase in pulmonary fibrosis. Antioxid. Redox Signal. 10, 343–354, doi: 10.1089/ars.2007.1908 (2008). PubMed DOI PMC

Schiller H. B. et al.. Time- and compartment-resolved proteome profiling of the extracellular niche in lung injury and repair. Mol. Syst. Biol. 11, 819, doi: 10.15252/msb.20156123 (2015). PubMed DOI PMC

Avissar N. et al.. Extracellular glutathione peroxidase in human lung epithelial lining fluid and in lung cells. Am. J. Physiol. 270, L173–182 (1996). PubMed

Matalon S., Holm B. A., Baker R. R., Whitfield M. K. & Freeman B. A. Characterization of antioxidant activities of pulmonary surfactant mixtures. Biochim. Biophys. Acta 1035, 121–127 (1990). PubMed

Park J. Y. et al.. Proteomic analysis of pancreatic juice for the identification of biomarkers of pancreatic cancer. J. Cancer Res. Clin. Oncol. 137, 1229–1238, doi: 10.1007/s00432-011-0992-2 (2011). PubMed DOI PMC

Naba A. et al.. The matrisome: in silico definition and in vivo characterization by proteomics of normal and tumor extracellular matrices. Mol. Cell. Proteomics 11, M111 014647, doi: 10.1074/mcp.M111.014647 (2012). PubMed DOI PMC

Rush J. W. E. & Sandiford S. D. Plasma glutathione peroxidase in healthy young adults: influence of gender and physical activity. Clin. Biochem. 36, 345–351, doi: 10.1016/S0009-9120(03)00039-0 (2003). PubMed DOI

Meplan C. et al.. Genetic polymorphisms in the human selenoprotein P gene determine the response of selenoprotein markers to selenium supplementation in a gender-specific manner (the SELGEN study). FASEB J. 21, 3063–3074, doi: 10.1096/fj.07-8166com (2007). PubMed DOI

Olivieri O. et al.. Selenium status, fatty acids, vitamins A and E, and aging: the Nove Study. Am J Clin Nutr. 60, 510–517 (1994). PubMed

Ray A. L. et al.. Low serum selenium and total carotenoids predict mortality among older women living in the community: the women’s health and aging studies. J Nutr. 136, 172–176 (2006). PubMed

Akbaraly N. T. et al.. Selenium and Mortality in the Elderly: Results from the EVA Study. Clin Chem. 51, 2117–2123, doi: 10.1373/clinchem.2005.055301 (2005). PubMed DOI

Bermudez O. et al.. Gli1 mediates lung cancer cell proliferation and Sonic Hedgehog-dependent mesenchymal cell activation. PLoS One. 8, e63226. doi: 10.1371/journal.pone.0063226 (2013). PubMed DOI PMC

Staab-Weijnitz C. A. et al.. FK506-binding Protein 10 is a Potential Novel Drug Target for Idiopathic Pulmonary Fibrosis. Am. J. Respir. Crit. Care Med. 192, 455–467, doi: 10.1164/rccm.201412-2233OC (2015). PubMed DOI PMC

Comhair S. A., Bhathena P. R., Farver C., Thunnissen F. B. & Erzurum S. C. Extracellular glutathione peroxidase induction in asthmatic lungs: evidence for redox regulation of expression in human airway epithelial cells. FASEB J. 15, 70–78, doi: 10.1096/fj.00-0085com (2001). PubMed DOI

Olson G. E. et al.. Extracellular glutathione peroxidase (Gpx3) binds specifically to basement membranes of mouse renal cortex tubule cells. Am. J. Physiol. Renal Physiol. 298, F1244–1253, doi: 10.1152/ajprenal.00662.2009 (2010). PubMed DOI PMC

Comhair S. A., Lewis M. J., Bhathena P. R., Hammel J. P. & Erzurum S. C. Increased glutathione and glutathione peroxidase in lungs of individuals with chronic beryllium disease. Am. J. Respir. Crit. Care Med. 159, 1824–1829, doi: 10.1164/ajrccm.159.6.9810044 (1999). PubMed DOI

Rushworth S. A. & MacEwan D. J. HO-1 underlies resistance of AML cells to TNF-induced apoptosis. Blood 111, 3793–3801, doi: 10.1182/blood-2007-07-104042 (2008). PubMed DOI

Rushworth S. A., Shah S. & MacEwan D. J. TNF mediates the sustained activation of Nrf2 in human monocytes. J. Immunol. 187, 702–707, doi: 10.4049/jimmunol.1004117 (2011). PubMed DOI

Song Y., Shi Y., Ao L. H., Harken A. H. & Meng X. Z. TLR4 mediates LPS-induced HO-1 expression in mouse liver: role of TNF-alpha and IL-1beta. World J. Gastroenterol. 9, 1799–1803 (2003). PubMed PMC

Terry C. M., Clikeman J. A., Hoidal J. R. & Callahan K. S. Effect of tumor necrosis factor-alpha and interleukin-1 alpha on heme oxygenase-1 expression in human endothelial cells. Am. J. Physiol. 274, H883–891 (1998). PubMed

Lin T. H. et al.. Upregulation of heme oxygenase-1 inhibits the maturation and mineralization of osteoblasts. J. Cell. Physiol. 222, 757–768, doi: 10.1002/jcp.22008 (2010). PubMed DOI

Lee Y. S. et al.. Dysregulation of adipose glutathione peroxidase 3 in obesity contributes to local and systemic oxidative stress. Mol. Endocrinol. 22, 2176–2189, doi: 10.1210/me.2008-0023 (2008). PubMed DOI PMC

Michaeloudes C., Chang P. J., Petrou M. & Chung K. F. Transforming growth factor-beta and nuclear factor E2-related factor 2 regulate antioxidant responses in airway smooth muscle cells: role in asthma. Am. J. Respir. Crit. Care Med. 184, 894–903, doi: 10.1164/rccm.201011-1780OC (2011). PubMed DOI PMC

Bakin A. V. et al.. Smad3-ATF3 signaling mediates TGF-beta suppression of genes encoding Phase II detoxifying proteins. Free Radical Biol. Med. 38, 375–387, doi: 10.1016/j.freeradbiomed.2004.10.033 (2005). PubMed DOI

Michaeloudes C., Sukkar M. B., Khorasani N. M., Bhavsar P. K. & Chung K. F. TGF-beta regulates Nox4, MnSOD and catalase expression, and IL-6 release in airway smooth muscle cells. Am. J. Physiol. Lung Cell Mol. Physiol. 300, L295–304, doi: 10.1152/ajplung.00134.2010 (2011). PubMed DOI PMC

Huang Y., Fang W., Wang Y., Yang W. & Xiong B. Transforming growth factor-beta1 induces glutathione peroxidase-1 and protects from H2O2-induced cell death in colon cancer cells via the Smad2/ERK1/2/HIF-1alpha pathway. Int. J. Mol. Med. 29, 906–912, doi: 10.3892/ijmm.2012.901 (2012). PubMed DOI

Comhair S. A., Thomassen M. J. & Erzurum S. C. Differential induction of extracellular glutathione peroxidase and nitric oxide synthase 2 in airways of healthy individuals exposed to 100% O(2) or cigarette smoke. Am. J. Respir. Cell Mol. Biol. 23, 350–354, doi: 10.1165/ajrcmb.23.3.4076 (2000). PubMed DOI

Voetsch B. et al.. Promoter polymorphisms in the plasma glutathione peroxidase (GPx-3) gene: a novel risk factor for arterial ischemic stroke among young adults and children. Stroke 38, 41–49, doi: 10.1161/01.STR.0000252027.53766.2b (2007). PubMed DOI PMC

John G. et al.. The composition of cigarette smoke determines inflammatory cell recruitment to the lung in COPD mouse models. Clin. Sci. (Lond.) 126, 207–221, doi: 10.1042/CS20130117 (2014). PubMed DOI PMC

Schaab C., Geiger T., Stoehr G., Cox J. & Mann M. Analysis of high accuracy, quantitative proteomics data in the MaxQB database. Mol. Cell. Proteomics 11, M111 014068, doi: 10.1074/mcp.M111.014068 (2012). PubMed DOI PMC

Raghu G. et al.. An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary fibrosis: evidence-based guidelines for diagnosis and management. Am. J. Respir. Crit. Care Med. 183, 788–824, doi: 10.1164/rccm.2009-040GL (2011). PubMed DOI PMC

Sterclova M., Paluch P., Skibova J. & Vasakova M. Influence of age on manifestation, VC and TLCO values, and bronchoalveolar lavage cell counts of sarcoidosis and extrinsic allergic alveolitis. Clin. Respir. J. 9, 39–44, doi: 10.1111/crj.12102 (2015). PubMed DOI

Costabel U. & Hunninghake G. W. ATS/ERS/WASOG statement on sarcoidosis. Sarcoidosis Statement Committee. American Thoracic Society. European Respiratory Society. World Association for Sarcoidosis and Other Granulomatous Disorders. Eur. Respir. J. 14, 735–737 (1999). PubMed

Schamberger A. C., Staab-Weijnitz C. A., Mise-Racek N. & Eickelberg O. Cigarette smoke alters primary human bronchial epithelial cell differentiation at the air-liquid interface. Sci. Rep. 5, 8163, doi: 10.1038/srep08163 (2015). PubMed DOI PMC

Hruz T. et al.. Genevestigator V3: A reference expression database for the meta-analysis of transcriptomes. Adv. Bioinformatics 420747, doi: 10.1155/2008/420747 (2008). PubMed DOI PMC

Carmagnol F., Sinet P. M. & Jerome H. Selenium-dependent and non-selenium-dependent glutathione peroxidases in human tissue extracts. Biochim. Biophys. Acta 759, 49–57 (1983). PubMed