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Protective Effects of Peroxiredoxin 6 in Pro-Inflammatory Response Model Using Raw 264.7 Macrophages

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Abstract

The aim of the work was to study effects of peroxiredoxin 6 (PRDX6), a recombinant antioxidant protein, on the level of pro-inflammatory responses of RAW 264.7 macrophages to endotoxin exposure. Addition of LPS to the RAW 264.7 cell culture medium expectedly increased production of TNF-α, and addition of PRDX6 led to a significant (15-20%) decrease in its production. The level of production of another pro-inflammatory cytokine, IL-1β, which was significantly activated by endotoxin, was completely normalized under the PRDX6 action. Moreover, addition of PRDX6 reduced production of reactive oxygen species (ROS) induced by endotoxin and also prevented overexpression of the iNos gene in the RAW 264.7 cells. The results showed that PRDX6 had a suppressive effect on the expression of Nrf-2 gene and production of the transcription factor NRF-2 during the first 6 h of cell cultivation. Addition of endotoxin caused activation of the NF-κB and SAPK/JNK signaling cascades, while in the presence of PRDX6, activity of these signaling cascades decreases. It is known that the pro-inflammatory response of cells caused by exposure to bacterial LPS leads to activation of apoptosis and elimination of the damaged cells. Our studies confirm this, since exposure to LPS led to activation of the expression of P53 gene, a marker of apoptosis. Peroxiredoxin 6 added within the first hours of the development of acute pro-inflammatory response suppressed the P53 gene expression, indicating protective effect of PRDX6 that reduced apoptosis in the RAW 264.7 macrophages.

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Abbreviations

LPS:

lipopolysaccharides

NF-κB:

nuclear factor-κB

PRDX6:

peroxiredoxin 6

ROS:

reactive oxygen species

TNF-α:

tumor necrosis factor alpha

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Acknowledgments

Equipment used in the work [Infinite 200 microplate reader (Tecan, Austria)] was provided by the Center for Collective Use of the Pushchino Research Center.

Funding

The work was supported by the Russian Science Foundation (grant no. 23-24-00041).

Author information

Authors and Affiliations

  1. Institute of Cell Biophysics, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    Svetlana B. Parfenyuk, Olga V. Glushkova, Mars G. Sharapov, Maksim O. Khrenov, Sergey M. Lunin, Anna A. Kuzekova, Elvira K. Mubarakshina, Tatyana V. Novoselova & Elena G. Novoselova

  2. Voronezh State University of Engineering Technologies, 394036, Voronezh, Russia

    Dmitrii A. Cherenkov

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  1. Svetlana B. Parfenyuk
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  10. Elena G. Novoselova
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Contributions

S.B.P., E.G.N., concept and supervision of the work; S.B.P., O.V.G., M.G.Sh., M.O.Kh., E.K.M., T.V.N., A.A.K., experimental work; S.B.P., E.G.N., S.M.L., O.V.G., discussion of the research results; S.B.P., E.G.N., writing the manuscript; S.M.L., D.A.Ch., O.V.G., M.G.Sh., editing the manuscript.

Corresponding author

Correspondence to Svetlana B. Parfenyuk.

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The authors declare no conflict of interest in financial or any other sphere. This article does not contain any studies involving human participants or animals performed by any of the authors.

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Parfenyuk, S.B., Glushkova, O.V., Sharapov, M.G. et al. Protective Effects of Peroxiredoxin 6 in Pro-Inflammatory Response Model Using Raw 264.7 Macrophages. Biochemistry Moscow 88, 1156–1164 (2023). https://doi.org/10.1134/S0006297923080096

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