title: Mitochondrial defects by intracellular calcium overload versus endothelial cold ischemia/reperfusion injury.
abstract: Questions as to the critical stress factor and primary targets of cold ischemia/reperfusion (CIR) injury were addressed by comparing mitochondrial defects caused by (1) CIR injury and (2) intracellular Ca2+ overload. CIR was simulated in transformed human umbilical vein endothelial cell cultures (tEC) by 8 h cold anoxia in University of Wisconsin solution and reoxygenation at 37 degrees C. Intracellular Ca2+ concentrations were changed by permeabilization of suspended cells with digitonin in culture medium (RPMI, 0.4 mM Ca2+). Binding of free Ca2+ by ethylene glycol-bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid in RPMI or mitochondrial incubation medium served as controls. Extracellular Ca2+ protected the cell membrane against permeabilization. Mitochondrial functions were determined before and after permeabilization of the cell membrane. After CIR, mitochondrial respiratory capacity declined, but oxygen consumption remained coupled to adenosine triphosphate (ATP) production. In contrast, Ca2+ overload caused uncoupling of mitochondrial respiration. High intracellular Ca2+ overload, therefore, does not reproduce cold ischemia/reperfusion injury in endothelial cells.
authors: Department of Transplant Surgery, University Hospital Innsbruck, Austria. erich.gnaiger@uibk.ac.at
authors: E
authors: E
authors: Gnaiger
authors: A V
authors: AV
authors: Kuznetsov
authors: G
authors: G
authors: Rieger
authors: A
authors: A
authors: Amberger
authors: A
authors: A
authors: Fuchs
authors: S
authors: S
authors: Stadlmann
authors: T
authors: T
authors: Eberl
authors: R
authors: R
authors: Margreiter
mesh_terms: D000241:Adenosine; D000493:Allopurinol; D002118:Calcium / Q000494:pharmacology / Q000502:physiology*; D015687:Cell Hypoxia; D002461:Cell Line, Transformed; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003080:Cold Temperature; D004533:Egtazic Acid; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D005978:Glutathione; D006801:Humans; D007328:Insulin; D007511:Ischemia; D008928:Mitochondria / Q000502:physiology*; D019188:Organ Preservation Solutions; D010101:Oxygen Consumption; D011887:Raffinose; D015427:Reperfusion Injury; D014471:Umbilical Veins
chemical_list: D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D004533:Egtazic Acid; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D002118:Calcium