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Consumption of redox energy by glutathione metabolism contributes to hypoxia/ reoxygenation-induced injury in astrocytes

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Abstract

The role of glutathione during ischemia/reperfusion is still a controversial issue. Glutathione should exert beneficial effects in the situation of ischemia/reperfusion due to its antioxidative potency. However, increasing survival time after transient ischemia and hypoxia has been reported for glutathione depleted cells. This work was aimed to analyse whether glutathione metabolism essentially contributes to redox energy failure and subsequent cell damage during ischemia/reperfusion. For this purpose, primary astrocyte rich cell cultures were subjected to 1 h hypoxia followed by up to 4 h reoxygenation in combination with substrate deprivation and glutathione depletion. The ability of the cells to reduce MTT was used to quantify the redox power of the cells. Inhibition of glutathione synthesis by L-buthionine-(S,R)-sulfoximine (BSO) caused depletion of cellular glutathione within 24 h and increase in MTT reduction by about 10% under normoxic conditions. Reoxygenation following 1 h of hypoxia was associated with decrease in MTT reduction which was enhanced by substrate deprivation. Glutathione depletion reduced hypoxia-induced decrease in MTT reduction. Three hours of substrate deprivation prior hypoxia resulted in lower levels of MTT reduction during reoxygenaton. Our data suggest that in situations of oxidative stress such as ischemia/reperfusion, glutathione metabolism may causes decrease of the cellular redox energy below a threshold level required for basic cellular functions finally resulting in cell injury.

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Authors and Affiliations

  1. Institute of Clinical Chemistry and Pathological Biochemistry, Department of Pathological Biochemistry, Medical Faculty, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany

    Petr Makarov, Ingrid Wiswedel, Wolfgang Augustin & Lorenz Schild

  2. Institute of Theoretical and Experimental Biophysics, Russian Academy of Science, 142290, Pushchino, Moscow region, Russia

    Petr Makarov

  3. Institute of Biometry and Medical Informatics, Magdeburg, Germany

    Siegfried Kropf

  4. Bereich Pathologische Biochemie der Medizinischen Fakultät der Otto-von-Guericke-Universität Magdeburg, Leipziger Str. 44, 39120, Magdeburg, Germany

    Lorenz Schild

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  1. Petr Makarov
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Correspondence to Lorenz Schild.

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Makarov, P., Kropf, S., Wiswedel, I. et al. Consumption of redox energy by glutathione metabolism contributes to hypoxia/ reoxygenation-induced injury in astrocytes. Mol Cell Biochem 286, 95–101 (2006). https://doi.org/10.1007/s11010-005-9098-y

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  • DOI: https://doi.org/10.1007/s11010-005-9098-y

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