Protective effects of novel derivatives of vitamin D 3 and lumisterol against UVB-induced damage in human keratinocytes involve activation of Nrf2 and p53 defense mechanisms

Anyamanee Chaiprasongsuk, Zorica Janjetovic, Tae Kang Kim, Stuart G. Jarrett, John A. D'Orazio, Michael F. Holick, Edith K.Y. Tang, Robert C. Tuckey, Uraiwan Panich, Wei Li, Andrzej T. Slominski

Research output: Contribution to journalArticlepeer-review

79 Scopus citations

Abstract

We tested whether novel CYP11A1-derived vitamin D 3 - and lumisterol-hydroxyderivatives, including 1,25(OH) 2 D 3 , 20(OH)D 3 , 1,20(OH) 2 D 3 , 20,23(OH) 2 D 3 , 1,20,23(OH) 3 D 3 , lumisterol, 20(OH)L 3 , 22(OH)L 3 , 20,22(OH) 2 L 3 , and 24(OH)L 3 , can protect against UVB-induced damage in human epidermal keratinocytes. Cells were treated with above compounds for 24 h, then subjected to UVB irradiation at UVB doses of 25, 50, 75, or 200 mJ/cm 2 , and then examined for oxidant formation, proliferation, DNA damage, and the expression of genes at the mRNA and protein levels. Oxidant formation and proliferation were determined by the DCFA-DA and MTS assays, respectively. DNA damage was assessed using the comet assay. Expression of antioxidative genes was evaluated by real-time RT-PCR analysis. Nuclear expression of CPD, phospho-p53, and Nrf2 as well as its target proteins including HO-1, CAT, and MnSOD, were assayed by immunofluorescence and western blotting. Treatment of cells with the above compounds at concentrations of 1 or 100 nM showed a dose-dependent reduction in oxidant formation. At 100 nM they inhibited the proliferation of cultured keratinocytes. When keratinocytes were irradiated with 50–200 mJ/cm 2 of UVB they also protected against DNA damage, and/or induced DNA repair by enhancing the repair of 6-4PP and attenuating CPD levels and the tail moment of comets. Treatment with test compounds increased expression of Nrf2-target genes involved in the antioxidant response including GR, HO-1, CAT, SOD1, and SOD2, with increased protein expression for HO-1, CAT, and MnSOD. The treatment also stimulated the phosphorylation of p53 at Ser-15, increased its concentration in the nucleus and enhanced Nrf2 translocation into the nucleus. In conclusion, pretreatment of keratinocytes with 1,25(OH) 2 D 3 or CYP11A1-derived vitamin D 3 - or lumisterol hydroxy-derivatives, protected them against UVB-induced damage via activation of the Nrf2-dependent antioxidant response and p53-phosphorylation, as well as by the induction of the DNA repair system. Thus, the new vitamin D 3 and lumisterol hydroxy-derivatives represent promising anti-photodamaging agents.

Original languageEnglish
Article number101206
JournalRedox Biology
Volume24
DOIs
StatePublished - Jun 2019

Bibliographical note

Funding Information:
The study was supported by NIH grants 1R01AR073004-01A1 and R01AR071189-01A1 and by a VA merit grant (no. 1I01BX004293-01A1) to ATS, United States.

Publisher Copyright:
© 2019

Keywords

  • Epidermal keratinocytes
  • Lumisterol (L )
  • Nuclear factor E2-related factor 2 (Nrf2)
  • Photoprotective effects
  • Ultraviolet B (UVB)
  • Vitamin D hydroxy-derivatives

ASJC Scopus subject areas

  • Organic Chemistry
  • Clinical Biochemistry

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