Abstract

The effect of bacterial endotoxin (lipopolysaccharides from Escherichia coli, LPS) on cellular metabolism and drug biotransformation was studied in precision-cut rat liver slices (PCLS). Xenobiotic metabolism by PCLS was assessed by measuring phase I (midazolam hydroxylation) and phase II (paracetamol conjugates) enzyme activities. Nitrites formation was used as an indirect way to assess LPS-mediated activation of nitric oxide synthase (iNOS, type 2). PCLS incubation with various LPS doses results in a dose-dependent formation of nitrites. Such a nitrite formation is decreased by dexamethasone. After incubation of PCLS for 24 h LPS addition did not increase the basal nitrite formation, indicating that cells are not responsive any more. Paracetamol conjugation was unaffected by LPS treatment but midazolam hydroxylation was reduced by more than 50%. Such a loss is not due to cell impairment since neither survival (LDH leakage) nor cellular metabolism (protein synthesis or ATP content) were modified by LPS. Indeed, under defined conditions, namely Williams' medium E and O-2/CO2 (95:5), PCLS maintained both ATP and GSH levels and the capacity of hepatocytes to synthesize proteins. In conclusion, the in vitro model of PCLS reproduces the inhibitory effect of LPS on a CYP3A-dependent activity, allowing a mechanistic approach to study cell-cell interactions. (C) 2002 Published by Elsevier Science Ltd.