Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Arsenite Acutely Decreases Nitric Oxide Production via the ROS-Protein Phosphatase 1-Endothelial Nitric Oxide Synthase-Thr497 Signaling Cascade

  • Seo, Jungwon (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Lee, Jee Young (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Sung, Min-Sun (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Byun, Catherine Jeonghae (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Cho, Du-Hyong (Department of Pharmacology, School of Medicine, Eulji University) ;
  • Lee, Hyeon-Ju (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Park, Jung-Hyun (Department of Molecular Medicine, School of Medicine, Ewha Womans University) ;
  • Cho, Ho-Seong (Biosafety Research Institute and College of Veterinary Medicine, Chonbuk National University) ;
  • Cho, Sung-Jin (Department of Biology, College of Natural Sciences, Chungbuk National University) ;
  • Jo, Inho (Department of Molecular Medicine, School of Medicine, Ewha Womans University)
  • Received : 2014.09.25
  • Accepted : 2014.10.22
  • Published : 2014.11.30
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Abstract

Chronic (>24 h) exposure of arsenite, an environmental toxicant, has shown the decreased nitric oxide (NO) production in endothelial cells (EC) by decreasing endothelial NO synthase (eNOS) expression and/or its phosphorylation at serine 1179 ($eNOS-Ser^{1179}$ in bovine sequence), which is associated with increased risk of vascular diseases. Here, we investigated the acute (<24 h) effect of arsenite on NO production using bovine aortic EC (BAEC). Arsenite acutely increased the phosphorylation of $eNOS-Thr^{497}$, but not of $eNOS-Ser^{116}$ or $eNOS-Ser^{1179}$, which was accompanied by decreased NO production. The level of eNOS expression was unaltered under this condition. Treatment with arsenite also induced reactive oxygen species (ROS) production, and pretreatment with a ROS scavenger N-acetyl-L-cysteine (NAC) completely reversed the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Although protein kinase C (PKC) and protein phosphatase 1 (PP1) were reported to be involved in $eNOS-Thr^{497}$ phosphorylation, treatment with PKC inhibitor, Ro318425, and overexpression of various PKC isoforms did not affect the arsenite-stimulated $eNOS-Thr^{497}$ phosphorylation. In contrast, treatment with PP1 inhibitor, calyculin A, mimicked the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Lastly, we found decreased cellular PP1 activity in arsenite-treated cells, which was reversed by NAC. Overall, our study demonstrates firstly that arsenite acutely decreases NO production at least in part by increasing $eNOS-Thr^{497}$ phosphorylation via ROS-PP1 signaling pathway, which provide the molecular mechanism underlying arsenite-induced increase in vascular disease.

Keywords

References

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