Rauen U, Reuters I, Fuchs A, de Groot H
Institut für Physiologische Chemie, Universitätsklinikum, Essen, Germany.
Hepatology. 1997 Aug;26(2):351-7. doi: 10.1002/hep.510260215.
We have previously shown that the injury to cultured liver endothelial cells during cold incubation in University of Wisconsin (UW) solution is energy-dependent and is mediated by reactive oxygen species. Here we demonstrate that this reactive oxygen-mediated injury is specific neither to endothelial cells nor to UW solution: cultured hepatocytes incubated in cold (4 degrees C) UW solution or histidine-tryptophan-ketoglutarate (HTK) solution were injured under normoxic conditions (loss of viability, 63% +/- 10% after 48 hours of cold incubation in UW solution and 82% +/- 11% after 24 hours of cold incubation in HTK solution), whereas hypoxia was protective (loss of viability, 29% +/- 12% [UW] and 13% +/- 3% [HTK] after the same cold incubation times). The injury under normoxic conditions was also largely decreased by adding either the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) or the flavonoid silibinin to the solutions, or by preincubating the cells with the iron chelator deferoxamine before the hypothermic incubation. Marked lipid peroxidation was observed during cold incubation in both preservation solutions. These results suggest that the injury to cultured hepatocytes during cold incubation in UW and HTK solutions is mediated by reactive oxygen species as is the injury to cultured liver endothelial cells.
我们之前已经表明，在威斯康星大学（UW）溶液中进行冷孵育时，培养的肝内皮细胞所受损伤是能量依赖性的，且由活性氧介导。在此我们证明，这种活性氧介导的损伤既不是内皮细胞特有的，也不是UW溶液特有的：在冷（4℃）UW溶液或组氨酸 - 色氨酸 - 酮戊二酸（HTK）溶液中孵育的培养肝细胞在常氧条件下会受到损伤（在UW溶液中冷孵育48小时后活力丧失63%±10%，在HTK溶液中冷孵育24小时后活力丧失82%±11%），而缺氧具有保护作用（在相同冷孵育时间后，UW溶液中活力丧失29%±12%，HTK溶液中活力丧失13%±3%）。通过向溶液中添加自旋捕获剂5,5 - 二甲基 - 1 - 吡咯啉N - 氧化物（DMPO）或类黄酮水飞蓟宾，或者在低温孵育前用铁螯合剂去铁胺对细胞进行预孵育，常氧条件下的损伤也会大幅降低。在两种保存溶液的冷孵育过程中均观察到明显的脂质过氧化。这些结果表明，在UW和HTK溶液中冷孵育时，培养肝细胞所受损伤与培养的肝内皮细胞所受损伤一样，是由活性氧介导的。
