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The Nitrone Spin Trap 5,5-Dimethyl-1-pyrroline N-oxide Affects Stress Response and Fate of Lipopolysaccharide-Primed RAW 264.7 Macrophage Cells

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Abstract

The nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) is commonly used to study free radicals. Due to its free radical trapping properties, DMPO is thought to reduce free radial-mediated oxidative damage and other related cellular responses. The purpose of this study was to assess the effect of DMPO on lipopolysaccharide (LPS)-induced inflammation, endoplasmic reticulum (ER) stress, and apoptosis in RAW 264.7 cells. The results showed that DMPO at 50 mM inhibited inducible nitric oxide synthase expression when added shortly after LPS treatment (≤3 h). Interestingly, DMPO increased anti-inflammatory heme oxygenase-1 (HO-1) expression and reversed LPS-induced decrease in HO-1 expression. LPS could increase cellular ER stress as indicated by C/EBP homologous protein (CHOP) induction; DMPO reduced LPS effect on CHOP expression. Unexpectedly, DMPO had a synergistic effect with LPS on increased caspase-3 activity. Overall, DMPO harbors multiple modulating effects but may induce apoptosis in LPS-stressed cells when given at 50 mM, an effective dose for its anti-inflammatory activity in vitro. Our data provide clues for further understanding of the nitrone spin trap with therapeutic potential.

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Acknowledgments

The project was supported by Award Number 5R00ES015415-04 to DCR from the National Institute of Environmental Health Sciences. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Environmental Health Sciences or the National Institutes of Health. DCR is an Affiliated Researcher of the Laboratory of Pharmacology and Toxicology at the NIEHS, USNIH, and DHHS.

Conflict of Interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Medicine, Section of Gastroenterology, The University of Chicago, Chicago, IL, 60637, USA

    Zili Zhai

  2. Laboratory of Experimental Medicine and Therapeutics, IMIBIO-SL, CONICET, San Luis, Argentina

    Sandra E. Gomez-Mejiba & Dario C. Ramirez

  3. Department of Biochemistry and Biological Sciences, School of Chemistry, Biochemistry and Pharmacy, National University of San Luis, San Luis, 5700, San Luis, Argentina

    Sandra E. Gomez-Mejiba & Dario C. Ramirez

Authors
  1. Zili Zhai
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  2. Sandra E. Gomez-Mejiba
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Correspondence to Zili Zhai or Dario C. Ramirez.

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Zhai, Z., Gomez-Mejiba, S.E. & Ramirez, D.C. The Nitrone Spin Trap 5,5-Dimethyl-1-pyrroline N-oxide Affects Stress Response and Fate of Lipopolysaccharide-Primed RAW 264.7 Macrophage Cells. Inflammation 36, 346–354 (2013). https://doi.org/10.1007/s10753-012-9552-4

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