Antioxidant responses and NRF2 in synergistic developmental toxicity of PAHs in zebrafish

Timme-Laragy, A.R., Van Tiem, L.A., Linney, E.A., and Di Giulio, R.T.
Toxicological sciences : an official journal of the Society of Toxicology   109(2): 217-227 (Journal)
Registered Authors
Di Giulio, Richard T., Linney, Elwood
PAH, NRF2, redox, ROS, embryonic development, glutathione
MeSH Terms
  • Animals
  • Antioxidants/metabolism*
  • Benzoflavones/toxicity
  • Buthionine Sulfoximine/toxicity
  • Drug Synergism
  • Embryo, Nonmammalian/abnormalities
  • Embryo, Nonmammalian/drug effects*
  • Embryonic Development/drug effects
  • Enzyme Inhibitors/toxicity
  • Gene Expression Regulation/drug effects*
  • Gene Knockdown Techniques
  • Glutathione/analysis
  • NF-E2-Related Factor 2/metabolism*
  • Oxidation-Reduction
  • Oxidative Stress
  • Pericardial Effusion
  • Polycyclic Aromatic Hydrocarbons/toxicity*
  • Up-Regulation
  • Zebrafish/embryology*
  • Zebrafish Proteins/metabolism*
  • beta-Naphthoflavone/toxicity
  • tert-Butylhydroperoxide/pharmacology
19233942 Full text @ Toxicol. Sci.
Early piscine life-stages are sensitive to polycyclic aromatic hydrocarbon (PAH) exposure, which can cause pericardial effusion and craniofacial malformations. We previously reported that certain combinations of PAHs cause synergistic developmental toxicity, as observed with co-exposure to the aryl hydrocarbon receptor (AHR) agonist beta-naphthoflavone (BNF) and cytochrome P4501A inhibitor alpha-naphthoflavone (ANF). Herein, we hypothesized that oxidative stress is a component of this toxicity. We examined induction of antioxidant genes in zebrafish embryos (Danio rerio) exposed to BNF or ANF individually, a BNF+ANF combination, and a pro-oxidant positive control, tert-butylhydroperoxide (tBOOH). We measured total glutathione, and attempted to modulate deformities using the glutathione synthesis inhibitor buthionine sulfoxamine (BSO) and increase glutathione pools with N-acetyl cysteine (NAC). In addition, we used a morpholino to knockdown expression of the antioxidant response element transcription factor NRF2 to determine if this would alter gene expression or increase deformity severity. BNF+ANF co-exposure significantly increased expressions of superoxide dismutase1 and2, glutathione peroxidase 1, pi class glutathione-s-transferase, and glutamate cysteine-ligase to a greater extent than tBOOH, BNF, or ANF alone. BSO pretreatment decreased some glutathione levels, but did not worsen deformities, nor did NAC diminish toxicity. Knockdown of NRF2 increased mortality following tBOOH challenge, prevented significant upregulation of antioxidant genes following both tBOOH and BNF+ANF exposures, and exacerbated BNF+ANF-related deformities. Collectively, these findings demonstrate that antioxidant responses are a component of PAH synergistic developmental toxicity, and that NRF2 is protective against prooxidant and PAH challenges during development.
Genes / Markers
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes