Difference between revisions of "Cervinkova 2007 Altern Lab Anim"
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{{Publication | {{Publication | ||
|title=Cervinkova Z, Lotkova H, | |title=Cervinkova Z, Lotkova H, Krivaikova P, Rousar T, Kucera O, Tichy L, Cervinka M, Drahota Z (2007) Evaluation of mitochondrial function in isolated rat hepatocytes and mitochondria during oxidative stress. Altern Lab Anim 35:353-61. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/17650955 PMID: 17650955 Open Access] | |||
|authors=Cervinkova Z, Lotkova H, Krivaikova P, Rousar T, Kucera O, Tichy L, Cervinka M, Drahota Z | |authors=Cervinkova Z, Lotkova H, Krivaikova P, Rousar T, Kucera O, Tichy L, Cervinka M, Drahota Z | ||
|year=2007 | |year=2007 | ||
|journal= Altern | |journal=Altern Lab Anim | ||
|abstract=The majority of toxic agents act either fully or partially via oxidative stress, the liver, specifically the mitochondria in hepatocytes, being the main target. Maintenance of mitochondrial function is essential for the survival and normal performance of hepatocytes, which have a high energy requirement. Therefore, greater understanding of the role of mitochondria in hepatocytes is of fundamental importance. Mitochondrial function can be analysed in several basic models: hepatocytes cultured in vitro; mitochondria in permeabilised hepatocytes; and isolated mitochondria. The aim of our study was to use all of these approaches to evaluate changes in mitochondria exposed in vitro to a potent non-specific peroxidating agent, tert-butylhydroperoxide (tBHP), which is known to induce oxidative stress. A decrease in the mitochondrial membrane potential (MMP) was observed in cultured hepatocytes treated with tBHP, as illustrated by a significant reduction in Rhodamine 123 accumulation and by a decrease in the fluorescence of the JC-1 molecular probe. Respiratory Complex I in the mitochondria of permeabilised hepatocytes showed high sensitivity to tBHP, as documented by high-resolution respirometry. This could be caused by the oxidation of NADH and NADPH by tBHP, followed by the disruption of mitochondrial calcium homeostasis, leading to the collapse of the MMP. A substantial decrease in the MMP, as determined by tetraphenylphosphonium ion-selective electrode measurements, also confirmed the dramatic impact of tBHP-induced oxidative stress on mitochondria. Swelling was observed in isolated mitochondria exposed to tBHP, which could be prevented by cyclosporin A, which is evidence for the role of mitochondrial permeability transition. Our results demonstrate that all of the above-mentioned models can be used for toxicity assessment, and the data obtained are complementary. | |abstract=The majority of toxic agents act either fully or partially via oxidative stress, the liver, specifically the mitochondria in hepatocytes, being the main target. Maintenance of mitochondrial function is essential for the survival and normal performance of hepatocytes, which have a high energy requirement. Therefore, greater understanding of the role of mitochondria in hepatocytes is of fundamental importance. Mitochondrial function can be analysed in several basic models: hepatocytes cultured ''in vitro''; mitochondria in permeabilised hepatocytes; and isolated mitochondria. The aim of our study was to use all of these approaches to evaluate changes in mitochondria exposed ''in vitro'' to a potent non-specific peroxidating agent, tert-butylhydroperoxide (tBHP), which is known to induce oxidative stress. A decrease in the mitochondrial membrane potential (MMP) was observed in cultured hepatocytes treated with tBHP, as illustrated by a significant reduction in Rhodamine 123 accumulation and by a decrease in the fluorescence of the JC-1 molecular probe. Respiratory Complex I in the mitochondria of permeabilised hepatocytes showed high sensitivity to tBHP, as documented by high-resolution respirometry. This could be caused by the oxidation of NADH and NADPH by tBHP, followed by the disruption of mitochondrial calcium homeostasis, leading to the collapse of the MMP. A substantial decrease in the MMP, as determined by tetraphenylphosphonium ion-selective electrode measurements, also confirmed the dramatic impact of tBHP-induced oxidative stress on mitochondria. Swelling was observed in isolated mitochondria exposed to tBHP, which could be prevented by cyclosporin A, which is evidence for the role of mitochondrial permeability transition. Our results demonstrate that all of the above-mentioned models can be used for toxicity assessment, and the data obtained are complementary. | ||
| | |mipnetlab=CZ Hradec Kralove Cervinkova Z, CZ Pardubice Rousar T | ||
|discipline=Environmental Physiology; Toxicology | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
|organism=Rat | |organism=Rat | ||
|tissues= | |tissues=Liver | ||
|preparations=Intact | |preparations=Intact cells, Permeabilized cells, Isolated mitochondria | ||
|injuries=Oxidative stress;RONS | |||
| | |||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
| | |discipline=Environmental Physiology; Toxicology | ||
}} | }} | ||
Latest revision as of 10:17, 24 March 2015
| Cervinkova Z, Lotkova H, Krivaikova P, Rousar T, Kucera O, Tichy L, Cervinka M, Drahota Z (2007) Evaluation of mitochondrial function in isolated rat hepatocytes and mitochondria during oxidative stress. Altern Lab Anim 35:353-61. |
Cervinkova Z, Lotkova H, Krivaikova P, Rousar T, Kucera O, Tichy L, Cervinka M, Drahota Z (2007) Altern Lab Anim
Abstract: The majority of toxic agents act either fully or partially via oxidative stress, the liver, specifically the mitochondria in hepatocytes, being the main target. Maintenance of mitochondrial function is essential for the survival and normal performance of hepatocytes, which have a high energy requirement. Therefore, greater understanding of the role of mitochondria in hepatocytes is of fundamental importance. Mitochondrial function can be analysed in several basic models: hepatocytes cultured in vitro; mitochondria in permeabilised hepatocytes; and isolated mitochondria. The aim of our study was to use all of these approaches to evaluate changes in mitochondria exposed in vitro to a potent non-specific peroxidating agent, tert-butylhydroperoxide (tBHP), which is known to induce oxidative stress. A decrease in the mitochondrial membrane potential (MMP) was observed in cultured hepatocytes treated with tBHP, as illustrated by a significant reduction in Rhodamine 123 accumulation and by a decrease in the fluorescence of the JC-1 molecular probe. Respiratory Complex I in the mitochondria of permeabilised hepatocytes showed high sensitivity to tBHP, as documented by high-resolution respirometry. This could be caused by the oxidation of NADH and NADPH by tBHP, followed by the disruption of mitochondrial calcium homeostasis, leading to the collapse of the MMP. A substantial decrease in the MMP, as determined by tetraphenylphosphonium ion-selective electrode measurements, also confirmed the dramatic impact of tBHP-induced oxidative stress on mitochondria. Swelling was observed in isolated mitochondria exposed to tBHP, which could be prevented by cyclosporin A, which is evidence for the role of mitochondrial permeability transition. Our results demonstrate that all of the above-mentioned models can be used for toxicity assessment, and the data obtained are complementary.
• O2k-Network Lab: CZ Hradec Kralove Cervinkova Z, CZ Pardubice Rousar T
Labels:
Stress:Oxidative stress;RONS Organism: Rat Tissue;cell: Liver Preparation: Intact cells, Permeabilized cells, Isolated mitochondria
HRR: Oxygraph-2k
