Reoxygenation after cold hypoxic storage of cultured precision-cut rat liver slices: effects on cellular metabolism and drug biotransformation.
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.
Rekka
E
E
Unité de Pharmacocinétique, Métabolisme, Nutrition et Toxicologie, Département de Sciences Pharmaceutiques, Université Catholique de Louvain, Louvain, Belgium.
Evdokimova
E
E
Eeckhaudt
S
S
Calderon
P B
PB
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001711:Biotransformation; D003080:Cold Temperature; D015925:Cryopreservation; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D004734:Energy Metabolism; D005978:Glutathione / Q000032:analysis; D008070:Lipopolysaccharides; D008099:Liver; D008297:Male; D008874:Midazolam / Q000737:chemistry; D010084:Oxidation-Reduction; D010100:Oxygen; D051381:Rats; D017208:Rats, Wistar; D013250:Steroid Hydroxylases / Q000032:analysis; D013997:Time Factors
D008070:Lipopolysaccharides; D000255:Adenosine Triphosphate; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; C465138:Cyp3a23-3a1 protein, rat; D051544:Cytochrome P-450 CYP3A; C022657:steroid hormone 6-beta-hydroxylase; D005978:Glutathione; D008874:Midazolam; D010100:Oxygen