Mimicking of glutathione peroxidase deficiency by exposition of JAR cells to increased level of synthetic hydroperoxide.
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.
Hallmann
A
A
Department of Pharmaceutical Biochemistry, Medical University of Gdańsk, Poland.
Milczarek
R
R
Szkatuła
M
M
Woźniak
M
M
Spodnik
J  H
J
Klimek
J
J
D000255:Adenosine Triphosphate / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects* / Q000378:metabolism; D048430:Cell Shape; D002822:Choriocarcinoma; D005609:Free Radicals / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000172:deficiency*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation; D008054:Lipid Peroxides / Q000737:chemistry; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism; D020122:tert-Butylhydroperoxide / Q000494:pharmacology*
D005609:Free Radicals; D008054:Lipid Peroxides; D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate; D020122:tert-Butylhydroperoxide; D007770:L-Lactate Dehydrogenase; D005979:Glutathione Peroxidase