Glutathione transferase omega 1 is required for the lipopolysaccharide-stimulated induction of NADPH oxidase 1 and the production of reactive oxygen species in macrophages

Deepthi Menon; Rebecca Coll; Luke A J O'Neill; Philip G Board

doi:10.1016/j.freeradbiomed.2014.05.020

Glutathione transferase omega 1 is required for the lipopolysaccharide-stimulated induction of NADPH oxidase 1 and the production of reactive oxygen species in macrophages

Free Radic Biol Med. 2014 Aug:73:318-27. doi: 10.1016/j.freeradbiomed.2014.05.020. Epub 2014 May 27.

Authors

Deepthi Menon¹, Rebecca Coll², Luke A J O'Neill², Philip G Board³

Affiliations

¹ Department of Molecular Biosciences, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2600, Australia.
² School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin 2, Ireland.
³ Department of Molecular Biosciences, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2600, Australia. Electronic address: Philip.Board@anu.edu.au.

PMID: 24873723
DOI: 10.1016/j.freeradbiomed.2014.05.020

Abstract

Bacterial lipopolysaccharide (LPS) stimulation of macrophages and inflammation via the Toll-like receptor 4 (TLR4) signaling pathway through NF-κΒ generates reactive oxygen species (ROS) and proinflammatory cytokines such as IL-1β, IL-6, and TNFα. Because glutathione transferase Omega 1-1 (GSTO1-1) can catalyze redox reactions such as the deglutathionylation of proteins and has also been implicated in the release of IL-1β we investigated its role in the development of LPS-mediated inflammation. Our data show that shRNA knockdown of GSTO1-1 in macrophage-like J774.1A cells blocks the expression of NADPH oxidase 1 and the generation of ROS after LPS stimulation. Similar results were obtained with a GSTO1-1 inhibitor. To maintain high ROS levels during an inflammatory response, LPS stimulation causes the suppression of enzymes such as catalase and glutathione peroxidase that protect against oxidative stress. The knockdown of GSTO1-1 also attenuates this response. Our data indicate that GSTO1-1 needs to be catalytically active and mediates its effects on the LPS/TLR4 inflammatory pathway upstream of NF-κΒ. These data suggest that GSTO1-1 is a novel target for anti-inflammatory intervention.

Keywords: Free radicals; Glutathione transferase Omega 1; LPS; NADPH oxidase 1; TLR4.

MeSH terms

Active Transport, Cell Nucleus
Adaptor Proteins, Signal Transducing
Animals
Carrier Proteins / antagonists & inhibitors
Carrier Proteins / genetics*
Cell Line
Cytoskeletal Proteins
Free Radicals
Glutathione Transferase / antagonists & inhibitors
Glutathione Transferase / genetics*
Heat-Shock Proteins / genetics
Inflammation / immunology
Inflammation / pathology
Interleukin-1beta / biosynthesis
Interleukin-6 / biosynthesis
Kelch-Like ECH-Associated Protein 1
Lipopolysaccharides
Macrophages / metabolism*
Mice
NADH, NADPH Oxidoreductases / biosynthesis*
NADPH Oxidase 1
NF-E2-Related Factor 2
NF-kappa B p50 Subunit / metabolism
Oxidation-Reduction
Oxidative Stress
PPAR gamma / biosynthesis
RNA Interference
RNA, Small Interfering
Reactive Oxygen Species / metabolism*
Toll-Like Receptor 4 / metabolism*
Transcription Factor RelA / metabolism
Tumor Necrosis Factor-alpha / biosynthesis

Substances

Adaptor Proteins, Signal Transducing
Carrier Proteins
Cytoskeletal Proteins
Free Radicals
Gsto1 protein, mouse
Heat-Shock Proteins
IL1B protein, mouse
Interleukin-1beta
Interleukin-6
Keap1 protein, mouse
Kelch-Like ECH-Associated Protein 1
Lipopolysaccharides
NF-E2-Related Factor 2
NF-kappa B p50 Subunit
Nfe2l2 protein, mouse
PPAR gamma
RNA, Small Interfering
Reactive Oxygen Species
Rela protein, mouse
Tlr4 protein, mouse
Toll-Like Receptor 4
Transcription Factor RelA
Tumor Necrosis Factor-alpha
NADH, NADPH Oxidoreductases
NADPH Oxidase 1
Glutathione Transferase