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Evaluation of mitochondrial function in isolated rat hepatocytes and mitochondria during oxidative stress

Publication at Central Library of Charles University, Faculty of Medicine in Hradec Králové |
2007

Abstract

The aim of our study was to evaluate mitochondrial changes in isolated rat hepatocytes and isolated liver mitochondria exposed in vitro to a potent non-specific peroxidating agent tert-butylhydroperoxide(tBHP).Decrease in mitochondrial membrane potential(MMP) in tBHP treated hepatocytes in culture was documented by significant reduction of Rhodamine 123 accumulation and by fluorescence microscopy using JC-1 as a fluorescence probe.Dramatic impact of oxidative stress induced by tBHP on mitochondria confirms also substantial decrease of MMP measured using tetraphenylphosphonium-selective electrode.Swelling of mitochondria,which was prevented by Cyclosporine A evidences the role of mitochondrial permeability transition:The high sensitivity of respiratory Complex I could be caused by oxidation on NADH and NADPH by tBHP with following disruption of mitochondrial calcium homeostasis.Increased mitochondrial calcium concentration stimulated formation of reactive oxygen species in mitochondria.