Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.

<h4>Rationale</h4>The underlying pathophysiology of bronchopulmonary dysplasia includes a macrophage-mediated host response orchestrated by anti-inflammatory peroxisome proliferator-activated receptor gamma (PPARγ) and anti-oxidant nuclear factor (erythroid-derived 2)-like 2 (Nrf2). Thes...

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Main Authors: Christopher R Nitkin, Tracey L Bonfield
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184469&type=printable
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author Christopher R Nitkin
Tracey L Bonfield
author_facet Christopher R Nitkin
Tracey L Bonfield
author_sort Christopher R Nitkin
collection DOAJ
description <h4>Rationale</h4>The underlying pathophysiology of bronchopulmonary dysplasia includes a macrophage-mediated host response orchestrated by anti-inflammatory peroxisome proliferator-activated receptor gamma (PPARγ) and anti-oxidant nuclear factor (erythroid-derived 2)-like 2 (Nrf2). These have not yet been studied in combination. This study tested the hypothesis that combined inflammatory and oxidative stressors would interact and change PPARγ- and Nrf2-regulated gene expression and antioxidant capacity. Therefore, we investigated the effect of dual stimulation with lipopolysaccharide and hyperoxia in murine bone marrow-derived macrophages (BMDM).<h4>Methods</h4>Sub-confluent BMDM from wild-type C57BL/6J mice were treated with lipopolysaccharide (LPS) 1ug/mL for 2 hours followed by room air (21% oxygen) or hyperoxia (95% oxygen) for 24 hours. Taqman real time-polymerase chain reaction gene expression assays, total antioxidant capacity assays, and Luminex assays were performed.<h4>Results</h4>Supernatants of cultured BMDM contained significant antioxidant capacity. In room air, LPS treatment decreased expression of PPARγ and Nrf2, and increased expression of tumor necrosis factor-alpha and heme oxygenase-1; similar findings were observed under hyperoxic conditions. LPS treatment decreased cellular total antioxidant capacity in room air but not in hyperoxia. Increased expression of sulfiredoxin-1 in response to hyperoxia was not observed in LPS-treated cells. Dual stimulation with LPS treatment and exposure to hyperoxia did not have synergistic effects on gene expression. Cellular total antioxidant capacity was not changed by hyperoxia exposure.<h4>Conclusions</h4>Our hypothesis was supported and we demonstrate an interaction between inflammatory and oxidative stressors in a model system of bronchopulmonary dysplasia pathogenesis. The protective anti-oxidant effect of cell culture media may have protected the cells from the most deleterious effects of hyperoxia.
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spelling doaj-art-589e93700a36497db9e61379b36762342025-08-20T02:45:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01129e018446910.1371/journal.pone.0184469Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.Christopher R NitkinTracey L Bonfield<h4>Rationale</h4>The underlying pathophysiology of bronchopulmonary dysplasia includes a macrophage-mediated host response orchestrated by anti-inflammatory peroxisome proliferator-activated receptor gamma (PPARγ) and anti-oxidant nuclear factor (erythroid-derived 2)-like 2 (Nrf2). These have not yet been studied in combination. This study tested the hypothesis that combined inflammatory and oxidative stressors would interact and change PPARγ- and Nrf2-regulated gene expression and antioxidant capacity. Therefore, we investigated the effect of dual stimulation with lipopolysaccharide and hyperoxia in murine bone marrow-derived macrophages (BMDM).<h4>Methods</h4>Sub-confluent BMDM from wild-type C57BL/6J mice were treated with lipopolysaccharide (LPS) 1ug/mL for 2 hours followed by room air (21% oxygen) or hyperoxia (95% oxygen) for 24 hours. Taqman real time-polymerase chain reaction gene expression assays, total antioxidant capacity assays, and Luminex assays were performed.<h4>Results</h4>Supernatants of cultured BMDM contained significant antioxidant capacity. In room air, LPS treatment decreased expression of PPARγ and Nrf2, and increased expression of tumor necrosis factor-alpha and heme oxygenase-1; similar findings were observed under hyperoxic conditions. LPS treatment decreased cellular total antioxidant capacity in room air but not in hyperoxia. Increased expression of sulfiredoxin-1 in response to hyperoxia was not observed in LPS-treated cells. Dual stimulation with LPS treatment and exposure to hyperoxia did not have synergistic effects on gene expression. Cellular total antioxidant capacity was not changed by hyperoxia exposure.<h4>Conclusions</h4>Our hypothesis was supported and we demonstrate an interaction between inflammatory and oxidative stressors in a model system of bronchopulmonary dysplasia pathogenesis. The protective anti-oxidant effect of cell culture media may have protected the cells from the most deleterious effects of hyperoxia.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184469&type=printable
spellingShingle Christopher R Nitkin
Tracey L Bonfield
Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
PLoS ONE
title Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
title_full Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
title_fullStr Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
title_full_unstemmed Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
title_short Balancing anti-inflammatory and anti-oxidant responses in murine bone marrow derived macrophages.
title_sort balancing anti inflammatory and anti oxidant responses in murine bone marrow derived macrophages
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184469&type=printable
work_keys_str_mv AT christopherrnitkin balancingantiinflammatoryandantioxidantresponsesinmurinebonemarrowderivedmacrophages
AT traceylbonfield balancingantiinflammatoryandantioxidantresponsesinmurinebonemarrowderivedmacrophages