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(The FASEB Journal. 1999;13:155-168.)
© 1999 FASEB

Research Communications

Cold-induced apoptosis in cultured hepatocytes and liver endothelial cells: mediation by reactive oxygen species

Ursula Rauena,1, Bernhard Polzarb, Harald Stephanb, Hans Georg Mannherzb and Herbert De Groota

a Institut für Physiologische Chemie, Universitätsklinikum, D-45122 Essen, Germany
b Abteilung für Anatomie und Embryologie, Ruhr-Universität, D-44780 Bochum, Germany

When cultured hepatocytes were incubated in cell culture medium at 4°C for up to 30 h and then returned to 37°C, blebbing of the plasma membrane, cell detachment, chromatin condensation and margination, enhanced nuclear stainability with Hoechst 33342, ruffling of the nuclear membrane, and DNA fragmentation occurred. Similar to hepatocytes, cultured liver endothelial cells exhibited blebbing, chromatin condensation and margination, marked nuclear condensation, and increased stainability with Hoechst 33342 when exposed to hypothermia/rewarming. In both cell types, the occurrence and extent of these alterations were dependent on the duration of the cold incubation period. This cold-induced apoptosis was inhibited by hypoxia, by an array of free radical scavengers/antioxidants, and by iron chelators. However, the extent of the protection by the different antioxidants was different in the two cell types: iron chelators provided complete protection in liver endothelial cells but only partial protection in hepatocytes, whereas lipophilic antioxidants such as {alpha}-tocopherol provided complete protection in both cell types. During cold incubation, and especially during rewarming, lipid peroxidation occurred. These results suggest that the formation of reactive oxygen species (ROS) is a key mediator of cold-induced apoptosis, with ROS formation being completely iron-mediated in liver endothelial cells and partially iron-mediated in hepatocytes.—Rauen, U., Polzar, B., Stephan, H., Mannherz, H. G., de Groot, H. Cold-induced apoptosis in cultured hepatocytes and liver endothelial cells: mediation by reactive oxygen species. FASEB J. 13, 155–168 (1999)


Key Words: hypothermia • iron • hydroxyl radical • free radical • cold preservation




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