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From bench to the hemodialysis clinic: protein-bound uremic toxins modulate NF-κB/Nrf2 expression

  • Nephrology - Original Paper
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Abstract

Purpose

Uremic toxins produced by gut microbiota (indoxyl sulfate—IS, p-cresyl sulfate—p-CS, and indole-3-acetic acid—IAA) accumulate in hemodialysis (HD) patients and exhibit potent inflammatory effects. However, the impact of these toxins on nuclear E2-related factor 2 (Nrf2) and nuclear factor-kappa B (NF-κB) expression in HD patients remains poorly defined. The aim of this study was to evaluate the association between uremic toxins and Nrf2/NF-κB expression in vitro (RAW 264.7 macrophage-like cells) and in peripheral blood mononuclear cells from HD patients.

Methods

Uremic toxins, C-reactive protein (CRP), interleukin-6 (IL-6) and malondialdehyde (MDA) levels were measured in fifteen HD patients and nine healthy individuals. RAW 264.7 macrophage-like cells were incubated with IS, as a prototype of protein-bound uremic toxin. Nrf2 and NF-κB expressions were analyzed by RT-qPCR.

Results

HD patients presented high levels of inflammatory markers, MDA and uremic toxins. In addition, they presented high NF-κB and low Nrf2 expression. Uremic toxins were positively correlated with NF-κB expression (IS, ρ = 0.58, p < 0.003; p-CS, ρ = 0.71, p < 0.001; IAA, ρ = 0.62, p < 0.001) and negatively with Nrf2 (IS, ρ = − 0.48, p = 0.01; p-CS, ρ = − 0.46, p < 0.02). Uremic toxins also exhibited positive correlations with CRP and MDA levels. Multivariate analysis revealed that p-CS is a determinant factor of NF-κB expression. In RAW 264.7 culture, NF-κB mRNA expression was stimulated by IS, while Nrf2 was downregulated.

Conclusions

Thus, uremic toxins may stimulate NF-κB mRNA and decrease Nrf2 expression in HD patients and, consequently, trigger inflammation and oxidative stress.

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Acknowledgements

This study was supported by Conselho Nacional de Pesquisa (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Clinical Research Unit (UPC) at Fluminense Federal University (UFF), Comité Français d´Evaluation de la Coopération Universitaire avec le Brésil (COFECUB), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Institut National de la Santé et de la Recherche Medicale (INSERM) and Institut National des Sciences Appliquées de Lyon (INSA-Lyon).

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Authors and Affiliations

  1. Graduate Program in Cardiovascular Sciences, Fluminense Federal University (UFF), Niterói, RJ, Brazil

    Milena B. Stockler-Pinto, Natália A. Borges, Ludmila F. M. F. Cardozo & Denise Mafra

  2. Graduate Program in Nutrition Sciences, Fluminense Federal University (UFF), Niterói, RJ, Brazil

    Milena B. Stockler-Pinto & Denise Mafra

  3. Univ-Lyon, CarMeN Laboratory, Inserm U1060, INSA Lyon, INRA U1397, Université Claude Bernard Lyon 1, Villeurbanne, France

    Christophe O. Soulage

  4. Departamento de Patologia Básica, Federal University of Paraná (UFPR), Curitiba, PR, Brazil

    Carla J. Dolenga & Lia S. Nakao

  5. Pontifícia Universidade Católica do Paraná, Curitiba, PR, Brazil

    Roberto Pecoits-Filho

  6. Department of Nephrology, Centre Hopitalier Lyon Sud, INSERM 1060, CENS, Université de Lyon, Lyon, France

    Denis Fouque

Authors
  1. Milena B. Stockler-Pinto
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Corresponding author

Correspondence to Milena B. Stockler-Pinto.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee of Medicine Faculty of Federal University Fluminense and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individuals participants included in the study.

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Stockler-Pinto, M.B., Soulage, C.O., Borges, N.A. et al. From bench to the hemodialysis clinic: protein-bound uremic toxins modulate NF-κB/Nrf2 expression. Int Urol Nephrol 50, 347–354 (2018). https://doi.org/10.1007/s11255-017-1748-y

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