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Use of pifithrin to inhibit p53-mediated signalling of TNF in dystrophic muscles of mdx mice

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Abstract

Tumour Necrosis Factor (TNF) plays a major role in exacerbating necrosis of dystrophic muscle; however, the precise molecular mechanism underlying this effect of TNF is unknown. This study investigates the role that p53 plays in TNF-mediated necrosis of dystrophic myofibres by inhibiting p53 using pifithrin-α and three pifithrin-β analogues. Tissue culture studies using C2C12 myoblasts established that pifithrin-α was toxic to differentiating myoblasts at concentrations greater than 10 μM. While non-toxic concentrations of pifithrin-α did not prevent the TNF-mediated inhibition of myoblast differentiation, Western blots indicated that nuclear levels of p53 were higher in TNF-treated myoblasts indicating that TNF does elevate p53. In contrast, in vivo studies in adult mdx mice showed that pifithrin-α significantly reduced myofibre necrosis that resulted from voluntary wheel running over 48 h. These results support the hypothesis that p53 plays some role in TNF-mediated necrosis of dystrophic muscle and present a potential new target for therapeutic interventions.

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Acknowledgements

This research was funded by a grant (MG/TS) from the National Health and Medical Research Council of Australia. We thank David Sassoon and Giovanna Marazzi (Paris) for suggesting the use of pifithrin-α for this study and Bijanka Gebski (UWA) for assistance in tissue culture studies. In addition, helpful discussions with Dr Trevor Payne (UWA) on selecting suitable pifithrin analogues were greatly appreciated.

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  1. School of Anatomy and Human Biology, University of Western Australia, 35 Stirling Hwy (M309), Crawley, WA, 6009, Australia

    Felicity J. Waters, Thea Shavlakadze & Miranda D. Grounds

  2. School of Biomedical, Biomolecular and Chemical Sciences, University of Western Australia, 35 Stirling Hwy (M309), Crawley, WA, 6009, Australia

    Matthew J. McIldowie & Matthew J. Piggott

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  1. Felicity J. Waters
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Correspondence to Miranda D. Grounds.

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Waters, F.J., Shavlakadze, T., McIldowie, M.J. et al. Use of pifithrin to inhibit p53-mediated signalling of TNF in dystrophic muscles of mdx mice. Mol Cell Biochem 337, 119–131 (2010). https://doi.org/10.1007/s11010-009-0291-2

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