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Caspase inhibition causes hyperacute tumor necrosis factor–induced shock via oxidative stress and phospholipase A2

Nature Immunology volume 4, pages 387–393 (2003)Cite this article

Abstract

Dysregulated apoptotic cell death contributes to many pathological conditions, including sepsis, prompting the suggestion that caspase inhibition to block apoptosis could have useful therapeutic applications. Because the cytokine tumor necrosis factor (TNF, also known as TNF-α) is both pro-apoptotic and pro-inflammatory and is involved in septic shock, we tested whether caspase inhibition would alleviate TNF-induced toxicity in vivo. General caspase inhibition by the protease inhibitor zVAD-fmk exacerbated TNF toxicity by enhancing oxidative stress and mitochondrial damage, resulting in hyperacute hemodynamic collapse, kidney failure and death. Thus, survival of TNF toxicity depends on caspase-dependent processes. Our results demonstrated the pathophysiological relevance of caspase-independent, ROS-mediated pathways in response to lethal TNF-induced shock in mice. In addition, survival of TNF toxicity seemed to require a caspase-dependent protective feedback on excessive reactive oxygen species (ROS) formation and phospholipase A2 activation.

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Figure 1: zVAD-fmk sensitizes mice to TNF but not to LPS.
Figure 2: zVAD-fmk sensitization and anti-oxidant treatment.
Figure 3: zVAD-fmk sensitization and PLA2, cyclooxygenase, 5-lipoxygenase or PAF inhibition.
Figure 4: zVAD-fmk sensitization reduces cPLA2 cleavage.
Figure 5: Cardiovascular analysis of zVAD-fmk sensitization.
Figure 6: HNE immunoreaction, H&E histology and electron microscopy.

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Acknowledgements

This research was supported by grants from the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen and the Interuniversitaire Attractiepolen. A.C. is a postdoctoral fellow with the FWO-Vlaanderen. We thank A. Raeymaekers and F. Duerinck for purifying TNF, E. Rogge and J. Hostens for help with immunoblotting and statistics, respectively, L. Van Geert, E. Spruyt and T. Hauttekeete for animal care, J. Debets for his preparation of mice in the hemodynamic studies and K. Vleminckx for critical reading of the manuscript.

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Authors and Affiliations

  1. Department for Molecular Biomedical Research, Molecular Pathophysiology and Experimental Therapy Unit, Ghent University and Flanders Interuniversity Institute for Biotechnology (VIB), K.L. Ledeganckstraat 35, Ghent, B-9000, Belgium

    Anje Cauwels & Peter Brouckaert

  2. Department of Pharmacology, Cardiovascular Research Institute Maastricht, University of Maastricht, Maastricht, 6200, MD, The Netherlands

    Ben Janssen

  3. Department of Pathology, Ghent University Hospital, Ghent, B-9000, Belgium

    Anouk Waeytens & Claude Cuvelier

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Correspondence to Anje Cauwels.

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Cauwels, A., Janssen, B., Waeytens, A. et al. Caspase inhibition causes hyperacute tumor necrosis factor–induced shock via oxidative stress and phospholipase A2. Nat Immunol 4, 387–393 (2003). https://doi.org/10.1038/ni914

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