Iron-dependent vs. iron-independent cold-induced injury to cultured rat hepatocytes: A comparative study in physiological media and organ preservation solutions☆
Section snippets
Animals
Male Wistar rats (250–300 g) were obtained from the Zentrales Tierlaboratorium (Universitätsklinikum Essen). Animals were kept under standard conditions with free access to food and water. All animals received humane care in compliance with the institutional guidelines.
Chemicals
Leibovitz L-15 medium, 2,2′-dipyridyl, choline chloride, choline bicarbonate, choline free base and lactobionic acid were obtained from Sigma–Aldrich (Taufkirchen, Germany). Deferoxamine mesylate was purchased from Novartis Pharma
Effect of iron chelators on cold-induced injury to rat hepatocytes during/after cold incubation in cell culture medium
When incubated for 24 h at 4 °C in cell culture medium (Leibovitz L-15) and rewarmed thereafter, cultured rat hepatocytes showed major cold-induced injury (Fig. 1). Part of this injury occurred during the cold incubation itself, part during rewarming. When cold incubation was performed in the presence of the iron chelator 2,2′-dipyridyl, cells were almost completely protected during cold incubation, but a major part of the protection was lost during rewarming. Practically identical results as
Discussion
We here further characterized cold-induced cell injury/apoptosis to rat hepatocytes and showed that the iron-independent component of cold-induced hepatocyte injury is a chloride-dependent component, predominantly occurs during rewarming, is associated with marked cell detachment and does not occur in organ preservation solutions.
Acknowledgments
We thank Mrs. E. Hillen and Ms. B. Lammers for their excellent technical assistance.
References (36)
- et al.
Hypothermia causes a marked injury to rat proximal tubular cells that is aggravated by all currently used preservation solutions
Cryobiology
(2003) - et al.
The role of DNA damage in the cytotoxic response to hydrogen peroxide/histidine
Gen. Pharmac.
(1997) - et al.
Selective role of intracellular chloride in the regulation of the intrinsic but not extrinsic pathway of apoptosis in Jurkat T-cells
J. Biol. Chem.
(2006) - et al.
Cold induces catalytic iron release of cytochrome P-450 origin: a critical step in cold storage-induced renal injury
Am. J. Transplant.
(2002) - et al.
Cold-induced apoptosis of rat liver cells in University of Wisconsin solution: the central role of chelatable iron
Hepatology
(2002) - et al.
Cold-induced release of reactive oxygen species as a decisive mediator of hypothermia injury to cultured liver cells
Free Radic. Biol. Med.
(1998) - et al.
Cold-induced apoptosis of hepatocytes: mitochondrial permeability transition triggered by nonmitochondrial chelatable iron
Free Radic. Biol. Med.
(2003) - et al.
Involvement of the mitochondrial pathway in cold storage and rewarming-associated apoptosis of human renal proximal tubular cells
Am. J. Transplant.
(2003) - et al.
Involvement of chloride in apoptotic cell death induced by activation of ATP-sensitive P2X7 purinoreceptor
J. Biol. Chem.
(2005) - et al.
Cold-induced apoptosis in isolated rat hepatocytes: protective role of glutathione
Free Radic. Biol. Med.
(2001)
Apoptosis vs. necrosis: glutathione-mediated cell death during rewarming of hepatocytes
Biochim. Biophys. Acta
Selective promotion of ferrous ion-dependent lipid peroxidation in Ehrlich ascites tumor cells by histidine as compared with other amino acids
Biochim. Biophys. Acta
Reoxygenation injury in rat hepatocytes: mediation by liberated by sources other than xanthine oxidase
Biol. Chem. Hoppe Seyler
A role for sodium in hypoxic but not in hypothermic injury to hepatocytes and LLC-PK1 cells
Transplantation
Rapid decrease in cellular sodium and chloride content during cold incubation of cultured liver endothelial cells and hepatocytes
Biochem. J.
Ion channels in cell proliferation and apoptotic cell death
J. Membr. Biol.
Experimental evaluation of Celsior, a new heart preservation solution
Eur. J. Cardiothorac. Surg.
Dual roles of plasmalemmal chloride channels in induction of cell death
Pflugers Arch.
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This study was supported by the Deutsche Forschungsgemeinschaft (RA 960/1-2 and RA 960/1-3).
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U.R. and H.d.G. are consultants of Dr. Franz Köhler Chemie GmbH, Alsbach-Hähnlein, Germany.