Rekka E, Evdokimova E, Eeckhaudt S, Calderon P B
Unité de Pharmacocinétique, Métabolisme, Nutrition et Toxicologie, Département de Sciences Pharmaceutiques, Université Catholique de Louvain, Louvain, Belgium.
Biochim Biophys Acta. 2001 Dec 19;1568(3):245-51. doi: 10.1016/s0304-4165(01)00225-2.
Cultured rat precision-cut liver slices (PCLS) were used to study the influence of hypothermic preservation and reoxygenation at 37 degrees C on cellular metabolism and drug biotransformation. Cold hypoxic storage caused a depressed metabolism in rat liver slices, but reoxygenation for 8 h at 37 degrees C partially restored the levels of both ATP and GSH and totally restored the capacity to synthesize proteins. Metabolism of midazolam (CYP3A-dependent oxidation) by cold preserved liver slices was decreased by 30% but no further affected by reoxygenation, showing the same profile as freshly cut slices. Such a reoxygenation at 37 degrees C is accompanied by a dramatic loss of CYP3A2 protein while CYP3A1 protein was unaffected. These results suggest that CYP3A2 did not play a major role in midazolam oxidation. Such results are not consistent with a putative reoxygenation injury but rather with cold hypoxic damage. Since cold preserved liver slices did not respond to bacterial endotoxin stimulation (lipopolysaccharides), a minor role of non-parenchymal cells is suggested as mediators for deleterious effects developed during the cold storage.
培养的大鼠精密肝切片（PCLS）用于研究低温保存及37℃复氧对细胞代谢和药物生物转化的影响。低温低氧储存导致大鼠肝切片代谢降低，但在37℃复氧8小时可部分恢复ATP和谷胱甘肽（GSH）水平，并完全恢复蛋白质合成能力。低温保存的肝切片对咪达唑仑（CYP3A依赖的氧化反应）的代谢降低了30%，但复氧未进一步影响，其表现与新鲜切片相同。37℃复氧伴随着CYP3A2蛋白的显著丢失，而CYP3A1蛋白未受影响。这些结果表明，CYP3A2在咪达唑仑氧化中不起主要作用。这些结果与假定的复氧损伤不一致，而与低温低氧损伤一致。由于低温保存的肝切片对细菌内毒素刺激（脂多糖）无反应，提示非实质细胞在低温储存期间产生的有害作用中起次要介导作用。
