Superoxide anion radical induced phototoxicity of 2,4,5,6-Tetraminopyrimidine sulfate via mitochondrial-mediated apoptosis in human skin keratinocytes at ambient UVR exposure

Food Chem Toxicol. 2022 Jun:164:112990. doi: 10.1016/j.fct.2022.112990. Epub 2022 Apr 6.

Abstract

2,4,5,6-Tetraaminopyrimidine sulfate (TAPS) is worldwide the most commonly used developer in hair dyes. As skin is the major organ, which is directly exposed to these permanent hair dyes, a comprehensive dermal safety assessment is needed. Hereto, we studied the photosensitization potential and mechanism involved in dermal phototoxicity of TAPS exposed to the dark and UVA/UVB/Sunlight by using different in-chemico and mammalian (HaCaT) cells, as test systems. Our experimental outcomes illustrate that TAPS get photodegraded (LC-MS/MS) and specifically generated superoxide anion radical (O2•-) under UVA and UVB via type-I photodynamic reaction. The phototoxic potential of TAPS is measured through MTT, NRU, and LDH assays that depicted a significant cell viability reduction at 25 μg/ml concentration and higher. Different cellular stainings (PI uptake, AO/EB, JC-1, NR uptake) suggested the role of mitochondrial-mediated apoptosis. Further, the transcriptomics study revealed upregulation of Apaf-1, Bax, Cytochrome c, Caspase 3, Caspase 9 and downregulation of Catalase and Bcl-2 by TAPS treated cells that strengthen our findings. Thus, the above findings suggest that chronic application of TAPS may be hazardous for human skin and promote various skin diseases.

Keywords: 2,4,5,6-Tetraaminopyrimidine sulfate; Apoptosis; DNA damage; Oxidative stress; Phototoxicity; Reactive oxygen species.

MeSH terms

  • Apoptosis
  • Chromatography, Liquid
  • DNA Damage
  • Dermatitis, Phototoxic* / metabolism
  • Hair Dyes*
  • Humans
  • Keratinocytes / metabolism
  • Reactive Oxygen Species / metabolism
  • Sulfates
  • Superoxides / metabolism
  • Tandem Mass Spectrometry
  • Ultraviolet Rays

Substances

  • Hair Dyes
  • Reactive Oxygen Species
  • Sulfates
  • Superoxides