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Glutathione depletion and increased apoptosis rate in human cystinotic proximal tubular cells

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Abstract

We have determined levels of glutathione (GSH), ATP, mitochondrial complex activity and apoptosis rate in proximal tubular cells (PTCs) exfoliated from urine in cystinotic (n=9) and control (n=9) children. Intracellular GSH was significantly depleted in cystinotic PTCs compared with controls (6.8 nmol GSH/mg protein vs 11.8 nmol GSH/mg protein; P<0.001), but there were no significant differences in mitochondrial complex activities or ATP levels under basal conditions. Cystinotic PTCs showed significantly increased apoptosis rate. After PTCs had been stressed by hypoxia, there was further depletion of GSH in cystinotic and control PTCs (2.4 nmol GSH/mg protein vs 7.2 nmol GSH/mg protein; P<0.001). Hypoxic stress led to increased complex I and complex IV activities in control but not in cystinotic PTCs. ATP levels were significantly reduced in cystinotic PTCs after hypoxic stress (12.2 nmol/mg protein vs 26.9 nmol/mg protein; P<0.001). GSH depletion occurs in this in vitro model of cystinotic PTCs, is exaggerated by hypoxic stress and may contribute to reduced ATP and failure to increase complex I/IV activities. Apoptotic rate is also increased, and these mechanisms may contribute to cellular dysfunction in cultured, human cystinotic PTCs.

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Acknowledgements

We thank Dr. Tom Smolenski for measurements of ATP. We are grateful to patients and parents for providing the urine samples and to Dr. Sue Rigden at Guy’s Hospital for providing access to her patients. This work was supported by the SDI-grant (Great Ormond Street Hospital, London, UK), the University Children’s Hospital Zurich (Switzerland) and the Swiss National Science Foundation (Berne/Zurich, Switzerland).

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Authors and Affiliations

  1. Nephro-urology Unit, Institute of Child Health, University College London Medical School, London, UK

    Guido F. Laube, Vanita Shah, Mushfequr R. Haq & William G. van’t Hoff

  2. Department of Molecular Neurosciences, Institute of Neurology, University College Medical School, London, UK

    Victoria C. Stewart, Iain P. Hargreaves & Simon J. R. Heales

  3. Neurometabolic Unit, National Hospital for Neurology and Neurosurgery, Queen Square, London, UK

    Iain P. Hargreaves & Simon J. R. Heales

  4. University Children’s Hospital, Steinwiesstrasse 75, 8032, Zurich, Switzerland

    Guido F. Laube

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  1. Guido F. Laube
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Correspondence to Guido F. Laube.

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Laube, G.F., Shah, V., Stewart, V.C. et al. Glutathione depletion and increased apoptosis rate in human cystinotic proximal tubular cells. Pediatr Nephrol 21, 503–509 (2006). https://doi.org/10.1007/s00467-006-0005-x

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  • DOI: https://doi.org/10.1007/s00467-006-0005-x

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