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Molecular Biology Reports

, Volume 46, Issue 6, pp 5785–5793 | Cite as

Superoxide-hydrogen peroxide imbalance differentially modulates the keratinocytes cell line (HaCaT) oxidative metabolism via Keap1-Nrf2 redox signaling pathway

  • Micheli Lamberti JobimEmail author
  • Verônica Farina Azzolin
  • Charles Elias Assmann
  • Vera Maria Melchiors Morsch
  • Ivana Beatrice Mânica da Cruz
  • Liliane de Freitas Bauermann
Original Article
  • 118 Downloads

Abstract

The purpose of this study was to investigate the effect of a superoxide-hydrogen peroxide (S-HP) imbalance of the superoxide dismutase manganese dependent (SOD2) gene, generated by paraquat and porphyrin exposure, on the keratinocytes cell line (HaCaT) oxidative metabolism. Paraquat acts increasing superoxide (O2·−) levels, while porphyrin increases hydrogen peroxide (H2O2) levels, acting as VV-SOD2-like and AA-SOD2-like molecules, respectively. First of all, HaCAT cells were treated with different concentrations of paraquat and porphyrin (1; 10; 30, and 70 μM) to determine the concentration of both that causes imbalance. After defining the concentration of paraquat and porphyrin (70 μM), a time curve was performed (1, 3, 6, and 24 h) to evaluate ROS production levels. Other oxidative parameters, such as nitric oxide (NO), lipoperoxidation (TBARS) and protein carbonyl, were evaluated after 24 h of incubation, as well as genotoxic analyses, apoptosis detection, and gene expression. Our findings revealed that paraquat exposure decreased cell viability, increasing lipoperoxidation, DNA damage, and apoptosis. On the other hand, porphyrin treatment increased cell viability and proliferation, ROS and NO production, triggering protein and DNA damage. In addition, porphyrin up-regulated Keap1 and Nrf2 gene expression, while paraquat decreased Nrf2 gene expression. In this sense, we suggested that the superoxide-hydrogen peroxide imbalance differentially modulates oxidative stress on keratinocytes cell line via Keap1-Nrf2 gene expression pathway.

Keywords

SOD2 Val16Ala-SOD2 SNP Paraquat Porphyrin Oxidative stress Keap1-Nrf2 pathway 

Notes

Funding

This study was supported by grants and fellowships from the following Brazilian governmental agencies: Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Micheli Lamberti Jobim
    • 1
    Email author
  • Verônica Farina Azzolin
    • 1
  • Charles Elias Assmann
    • 2
  • Vera Maria Melchiors Morsch
    • 2
  • Ivana Beatrice Mânica da Cruz
    • 1
    • 3
  • Liliane de Freitas Bauermann
    • 1
  1. 1.Programa de Pós-graduação em FarmacologiaUniversidade Federal de Santa MariaSanta MariaBrazil
  2. 2.Programa de Pós-graduação em Ciências Biológicas: Bioquímica ToxicológicaUniversidade Federal de Santa MariaSanta MariaBrazil
  3. 3.Programa de Pós-graduação em GerontologiaUniversidade Federal de Santa MariaSanta MariaBrazil

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