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Glutathione peroxidase 3 localizes to the epithelial lining fluid and the extracellular matrix in lung fibrosis

Andrea Schamberger, Herbert Schiller, Isis Fernandez, Martina Sterclova, Katharina Heinzelmann, Elisabeth Hennen, Rudolf Hatz, Jürgen Behr, Martina Vasakova, Matthias Mann, Oliver Eickelberg, Claudia Staab-Weijnitz
European Respiratory Journal 2016 48: PA781; DOI: 10.1183/13993003.congress-2016.PA781
Andrea Schamberger
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Herbert Schiller
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Isis Fernandez
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Martina Sterclova
2Department of Pneumology, Thomayer Hospital, Prague 4-Krc, Czech Republic
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Katharina Heinzelmann
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Elisabeth Hennen
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Rudolf Hatz
3Thoraxchirurgisches Zentrum, Klinik für Allgemeine-, Viszeral-, Transplantations-, Gefäß- und Thoraxchirurgie, University Hospital of the Ludwig-Maximilians University Munich, Munich, Germany
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Jürgen Behr
4Medizinische Klinik und Poliklinik V, Member of the German Center for Lung Research (DZL), University Hospital of the Ludwig-Maximilians University Munich, Munich, Germany
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Martina Vasakova
2Department of Pneumology, Thomayer Hospital, Prague 4-Krc, Czech Republic
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Matthias Mann
5Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany
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Oliver Eickelberg
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Claudia Staab-Weijnitz
1Comprehensive Pneumology Center, Member of the German Center for Lung Research (DZL), Helmholtz Zentrum München, Munich, Germany
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Abstract

Aberrant antioxidant activity contributes to the pathogenesis of interstitial lung diseases (ILD). The composition of the extracellular matrix (ECM) is affected by oxidative stress, but extracellular antioxidant defense mechanisms in ILD are incompletely understood.

Here, relative abundance and detergent solubility of extracellular antioxidant enzymes were extracted from a proteomic dataset of bleomycin-induced lung fibrosis in C57BL/6N mice. Glutathione peroxidase (Gpx) activity was measured in bronchoalveolar lavage fluid (BALF) and immunofluorescent stainings were performed to address distribution of Gpx3 in the lung. Regulation of GPX3 by TGF-β1, TNF-α, and menadione was assessed in primary human bronchial epithelial cells (phBEC) and lung fibroblasts (phLF) by qPCR and ELISA. Finally, GPX3 levels were measured in BALF from ILD patients by ELISA.

Sod3, Gpx3, and Gpx activity were increased in BALF of mice subjected to bleomycin-induced lung fibrosis. In total lung tissue, only Gpx3, but not Sod3, was upregulated. Quantitative detergent solubility profiles indicated that Gpx3, but not Sod3, 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 mouse and human lung tissue. GPX3 expression in phBEC and phLF was downregulated by TNF-α, but more variably regulated by TGF-β1 and menadione. BALF of some ILD patients contained high levels of GPX3.

Hence, levels of the extracellular antioxidant enzyme GPX3 are variably increased in ILD. Upregulation of GPX3 might protect the ECM from oxidative damage in ILD.

  • Interstitial lung disease
  • Proteomics
  • Bronchoalveolar lavage
  • Copyright ©the authors 2016
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Glutathione peroxidase 3 localizes to the epithelial lining fluid and the extracellular matrix in lung fibrosis
Andrea Schamberger, Herbert Schiller, Isis Fernandez, Martina Sterclova, Katharina Heinzelmann, Elisabeth Hennen, Rudolf Hatz, Jürgen Behr, Martina Vasakova, Matthias Mann, Oliver Eickelberg, Claudia Staab-Weijnitz
European Respiratory Journal Sep 2016, 48 (suppl 60) PA781; DOI: 10.1183/13993003.congress-2016.PA781

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Glutathione peroxidase 3 localizes to the epithelial lining fluid and the extracellular matrix in lung fibrosis
Andrea Schamberger, Herbert Schiller, Isis Fernandez, Martina Sterclova, Katharina Heinzelmann, Elisabeth Hennen, Rudolf Hatz, Jürgen Behr, Martina Vasakova, Matthias Mann, Oliver Eickelberg, Claudia Staab-Weijnitz
European Respiratory Journal Sep 2016, 48 (suppl 60) PA781; DOI: 10.1183/13993003.congress-2016.PA781
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