1. Mimicking of glutathione peroxidase deficiency by exposition of JAR cells to increased level of synthetic hydroperoxide.
- Author
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Hallmann A, Milczarek R, Szkatuła M, Woźniak M, Spodnik J, and Klimek J
- Subjects
- Adenosine Triphosphate metabolism, Cell Line, Tumor metabolism, Cell Shape, Choriocarcinoma, Free Radicals metabolism, Humans, L-Lactate Dehydrogenase metabolism, Lipid Peroxidation, Lipid Peroxides chemistry, Mitochondria metabolism, Mitochondria ultrastructure, Oxidation-Reduction, Reactive Oxygen Species metabolism, Cell Line, Tumor drug effects, Glutathione Peroxidase deficiency, tert-Butylhydroperoxide pharmacology
- Abstract
A short chain synthetic analogue of lipid hydroperoxides was used to overload glutathione peroxidase (GPx) in human choriocarcinoma cell line JAR cells. Cells exposed to 100 microM tBuOOH displayed a 40% reduction in ATP level and significantly increased in membrane permeability, visualised by the lactate dehydrogenase (LDH) release into the extracellular medium. The intracellular level of oxygen free radicals measured as an oxidation of the dichlorodihydro-fluorescein diacetate (H2DCF-DA) significantly increased after 2 hours of cell exposition to 100 microM tBuOOH. Concomitantly MDA, 4-HNE level increased to 2 nmol/mg of cell protein after 2 hours. Mitochondria stained with MitoTracker Red CMXRos displayed a filamentous appearance in control cells but changed into granular less energised organelles after exposition to tBuOOH. Collectively, the above results indicate the importance of the contribution of oxidative stress in the development of pre-eclampsia.
- Published
- 2005