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Spironolactone induces apoptosis and inhibits NF-κB independent of the mineralocorticoid receptor

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Abstract

Spironolactone (SPIR) binds to cytoplasmic mineralocorticoid receptors (MR) and functions as an aldosterone (ALDO) antagonist. Recently, however, the drug was shown to have an early MR independent, suppressive effect on immunoactive and inflammatory cytokines as well as an apoptotic effect on blood mononuclear cells (MNC). To elucidate the mechanism behind SPIR’s apoptotic effect, we investigated the relation between apoptosis and cytokine suppression for SPIR along with the apoptosis-inducing and antiinflammatory drug sulfasalazine (SFZ). Using human MNC, we found that SPIR and SFZ, at concentrations 10 and 1000 μM, respectively, significantly increased both apoptosis and cell death. Production of inflammatory cytokines was significantly reduced by 3 to 30 μM SPIR and by 300 to 1000 μM SFZ. We also found that 0.4 μM SPIR and 300 μM SFZ significantly reduced the activity of NF-κB, a transcription factor involved in both apoptosis and immunoinflammation. ALDO, the MR antagonist, eplerenone, and the SPIR metabolite, 7α-thiomethyl-spironolactone, slightly reduced NF-κB activity, but they did not interfere with SPIR’s effect, showing that MR binding is not involved in SPIR-induced suppression of NF-κB activity. Finally, phosphorylation of IκBα was also significantly reduced by SPIR. These results provide new insight into the apoptotic and anti-inflammatory effects of SPIR.

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Abbreviations

7TS:

7α-thiomethyl-spironolactone

ALDO:

Aldosterone

CD:

Crohn’s disease

EPL:

Eplerenone

GM-CSF:

Granulocyte-macrophage colony-stimulating factor

IFN:

Interferon

IL:

Interleukin

IkBα:

Inhibitor of κ light chain gene enhancer in B cells

LPS:

Lipopolysaccharide

MNC:

Mononuclear cells

MR:

Mineralocorticoid receptor

NF-κB:

Nuclear factor κB

PBS:

Phosphate buffered saline

PHA:

Phytohaemagglutinin-A

PI:

Propidium iodide

RA:

Rheumatoid arthritis

SFZ:

Sulfasalazine

SPIR:

Spironolactone

TNF:

Tumour necrosis factor

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Authors and Affiliations

  1. Institute for Inflammation Research, IIR7521, Rigshospitalet National University Hospital, Blegdamsvej, 9, DK-2100, Copenhagen E, Denmark

    Søren Ulrik Salling Sønder & Klaus Bendtzen

  2. Institute of Molecular Biology and Physiology, Department of Immunology and Institute of Medical Microbiology and Immunology, University of Copenhagen, Blegdamsvej 3, 2200, Copenhagen N, Denmark

    Anders Woetmann & Niels Ødum

Authors
  1. Søren Ulrik Salling Sønder
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  2. Anders Woetmann
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  3. Niels Ødum
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  4. Klaus Bendtzen
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Corresponding author

Correspondence to Klaus Bendtzen.

Additional information

Financial support: The Danish Research Agencies, the Danish Rheumatism Association, the Carla Thiel Foundation and the Danish Biotechnology Program.

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Sønder, S.U.S., Woetmann, A., Ødum, N. et al. Spironolactone induces apoptosis and inhibits NF-κB independent of the mineralocorticoid receptor. Apoptosis 11, 2159–2165 (2006). https://doi.org/10.1007/s10495-006-0286-3

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