ROS production and glutathione response in keratinocytes after application of β-carotene and VIS/NIR irradiation

Chem Biol Interact. 2018 Jan 25:280:1-7. doi: 10.1016/j.cbi.2017.12.002. Epub 2017 Dec 5.

Abstract

The skin is exposed to many stress factors which, in turn, can promote a shift of the antioxidant (AO) network towards the prooxidative side, supporting the development of various skin disorders. A balanced diet, in combination with a healthy lifestyle could reduce oxidative stress. Carotenoids are essential nonenzymatic AOs and main components of the exogenous AO system. To examine the interdependence between endogenous and exogenous AOs, secondary keratinocytes (HaCaT) were treated with various Beta (β-)-carotene concentrations with subsequent stress treatment by moderate irradiation (700-2000 nm). To facilitate the uptake of β-carotene, an innovative nanocrystal solution was used. Cell viability assay was applied to HaCaT cells to evaluate suitable concentration of β-carotene, whereby the uptake was measured by resonant Raman spectroscopy. The redox status was determined before and after supplementation with two selected β-carotene concentrations (0.02 and 0.1 μg/ml) and irradiation. Reactive oxygen species (ROS) were measured by electron paramagnetic resonance spectroscopy and the AO glutathione (GSH) by a fluorescent-based assay for evaluating the endogenous redox status. An increase of ROS and a reduction of GSH after irradiation was observed. Interestingly, the applied β-carotene, already induce oxidative stress. Nevertheless, an effective protection against irradiation could be observed for the lower dose. The high dose turned pro-oxidative.

Keywords: Antioxidants; Electron paramagnetic resonance (EPR) spectroscopy; Oxidative stress; VIS/NIR irradiation; β-carotene.

MeSH terms

  • Cell Line
  • Cell Survival / drug effects
  • Electron Spin Resonance Spectroscopy
  • Glutathione / metabolism*
  • Humans
  • Infrared Rays*
  • Keratinocytes / drug effects
  • Keratinocytes / metabolism
  • Keratinocytes / radiation effects
  • Light*
  • Oxidation-Reduction
  • Oxidative Stress / drug effects*
  • Oxidative Stress / radiation effects
  • Reactive Oxygen Species / metabolism*
  • Spectrum Analysis, Raman
  • beta Carotene / pharmacology*

Substances

  • Reactive Oxygen Species
  • beta Carotene
  • Glutathione