Reduced Nrf2 activation in PI3K phosphorylation-impaired vitiliginous keratinocytes increases susceptibility to ROS-generating chemical-induced apoptosis

Environ Toxicol. 2017 Dec;32(12):2481-2491. doi: 10.1002/tox.22461. Epub 2017 Aug 24.


Keratinocytes in affected epidermis of vitiligo patients are known to have impaired activation of the PI3K/AKT pathway. Based on critical roles of keratinocytes and oxidative stress in vitiligo development, this study examined whether keratinocytes with impaired PI3K activation were more vulnerable to apoptosis caused by oxidative stress from phenolic compounds, p-tert-butylphenol (4-TBP) and hydroquinone (HQ). Cell viability assay, FACS analysis, ELISA for TNF-α or IL-1a, ROS assay, Western blot analysis for Nrf2 expression, and confocal microscopy with anti-Nrf2 and phospho-PI3K antibodies were done on primary cultured normal human keratinocytes with or without PI3K knockdown in the presence or absence of chemical treatment or antioxidant. Immunofluorescence staining using anti-Nrf2, phospho-PI3K, TNF-ɑ, and IL-1ɑ antibodies, ROS assay using dihydroethidium, and TUNEL assay were performed on sets of depigmented and normally pigmented skin from vitiligo patients. Results showed that 4-TBP or HQ treatment increased apoptosis and the expression levels of TNF-ɑ, IL-1ɑ, and ROS in PI3K-knockdown keratinocytes which reduced Nrf2 nuclear translocation compared to control keratinocytes. These changes were significantly recovered by an antioxidant treatment. Depigmented epidermis of vitiligo patients also showed lower levels of Nrf2 and phospho-PI3K but higher levels of ROS, TNF-ɑ, IL-1ɑ, and ROS with more TUNEL-positive cells. Therefore, impaired PI3K activation in keratinocytes in depigmented epidermis of vitiligo patients are vulnerable to apoptosis caused by ROS-generating chemicals due to reduced Nrf2 activation.

Keywords: ROS-generating chemicals; apoptosis; impaired PI3K activation; reduced Nrf2 activation; vitiliginous keratinocytes.

MeSH terms

  • Adolescent
  • Adult
  • Antioxidants / pharmacology
  • Apoptosis*
  • Cell Survival / drug effects
  • Cells, Cultured
  • Epidermis / pathology
  • Female
  • Gene Knockdown Techniques
  • Humans
  • Hydroquinones / toxicity*
  • Interleukin-1alpha / metabolism
  • Keratinocytes / drug effects*
  • Keratinocytes / metabolism
  • Keratinocytes / pathology
  • Male
  • Melanocytes / drug effects
  • Melanocytes / pathology
  • Middle Aged
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidative Stress / drug effects
  • Phenols / toxicity*
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation
  • Reactive Oxygen Species / metabolism*
  • Tumor Necrosis Factor-alpha / metabolism
  • Vitiligo / metabolism*
  • Vitiligo / pathology
  • Young Adult


  • Antioxidants
  • Hydroquinones
  • Interleukin-1alpha
  • NF-E2-Related Factor 2
  • Phenols
  • Reactive Oxygen Species
  • Tumor Necrosis Factor-alpha
  • Phosphatidylinositol 3-Kinases
  • butylphen
  • hydroquinone