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Sulforaphane suppresses LPS-induced inflammation in primary rat microglia

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Abstract

Objective and design

The aim of this study was to investigate the signal transduction pathways involved in sulforaphane (SF) mediated inhibition of the inflammatory response to lipopolysaccharide (LPS). Additionally, we investigated the effects of SF and LPS on the activity of Nrf2.

Material

Primary rat microglia and the murine microglia cell line BV2 were used.

Treatment

Cells were treated with LPS with or without SF.

Methods

Cell viability was measured via WST-assay. Real-time RT-PCR was performed to analyze cytokine mRNA levels. The nitric oxide (NO) release was measured in LPS-stimulated microglia. The induction of various signal transduction pathways and Nrf2 was determined by Western blotting. NF-κB and AP-1 activation was measured by dual luciferase assay.

Results

We showed that SF attenuates the LPS-induced increase of IL-1β, IL-6, and TNF-α expression in microglia. In addition, SF significantly decreases the NO in a concentration-dependent manner. SF inhibits LPS-stimulated ERK1/2 and JNK phosphorylation and thereby inhibits the LPS-induced activation of NF-κB- and activator protein-1 (AP-1). Moreover, SF and LPS together are able to induce Nrf2 activation.

Conclusions

We showed that SF, and also LPS by itself, are able to activate the cell’s defence against oxidative and electrophilic stress. We conclude that SF could be a candidate agent for anti-inflammatory treatment of the central nervous system.

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Abbreviations

Nrf2:

Nuclear factor erythroid 2-related factor 2

DMEM:

Dulbecco’s modified Eagle’s medium

FCS:

Fetal calf serum

SFM:

Serum free medium

WST:

4-[3-(4-Iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1, 3-benzene disulfonate

p-:

Phospho-

ERK:

Extracellular signal-regulated kinase

ANOVA:

Analysis of variance

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Acknowledgments

We would like to thank Rosemarie Sprang, Christiane Jaeschke, Susanne Echterhagen, and Regine Worm for their excellent technical assistance. This study was supported by the “Forschungsförderung” of the medical faculty (University of Kiel, Germany, to L.O.B.), Hensel Foundation (University of Kiel, Germany, to L.O.B.), Deutsche Forschungsgemeinschaft (DFG PU214/5-2; PU214/4-2; PU214/3-2) and the Hertie Foundation (M.K.).

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, RWTH Aachen University, Wendlingweg 2, 52074, Aachen, Germany

    Lars-Ove Brandenburg, Thomas Pufe & Christoph J. Wruck

  2. Department of Anatomy, Christian-Albrechts-University Kiel, Kiel, Germany

    Lars-Ove Brandenburg & Ralph Lucius

  3. Department of Neuroanatomy, RWTH Aachen University, 52074, Aachen, Germany

    Markus Kipp

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  1. Lars-Ove Brandenburg
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  2. Markus Kipp
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  3. Ralph Lucius
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  4. Thomas Pufe
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  5. Christoph J. Wruck
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Corresponding author

Correspondence to Lars-Ove Brandenburg.

Additional information

Responsible Editor: L. Li.

T. Pufe and C. J. Wruck contributed equally to this article.

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Brandenburg, LO., Kipp, M., Lucius, R. et al. Sulforaphane suppresses LPS-induced inflammation in primary rat microglia. Inflamm. Res. 59, 443–450 (2010). https://doi.org/10.1007/s00011-009-0116-5

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  • DOI: https://doi.org/10.1007/s00011-009-0116-5

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