Comparative effects of rifabutin and rifampicin on cytochromes P450 and UDP-glucuronosyl-transferases expression in fresh and cryopreserved human hepatocytes

Chem Biol Interact. 1999 Jun 1;121(1):37-48. doi: 10.1016/s0009-2797(99)00089-7.


The aim of this study was to evaluate rifabutin (RBT) and rifampicin (RIF) capabilities in inducing various xenobiotic metabolizing enzymes such as cytochromes P450 (CYPs) and UDP-glucuronosyl-transferases (UGTs) in cultured fresh and cryopreserved human hepatocytes. Enzyme induction was assessed through the use of several diagnostic markers, i.e. testosterone, midazolam (MDZ), diazepam (DZP) and 7-ethoxyresorufin for CYP-dependent enzyme reactions; and AZT for UGT-dependent enzyme reactions. RBT concentrations (0.118, 0.708 microM) were selected according to previously published pharmacokinetic data in patients. The known CYP3A4 inducer in humans, RIF, was used as a positive control. At the concentrations used, no sign of cytotoxicity was evidenced. Both compounds were able to dose-dependently induce the overall metabolism of testosterone (approximately 2-fold for RBT, 4-fold for RIF) and the formation of the 6beta-hydroxylated-derivative (up to approximately 4-fold over control for RBT and approximately 10-fold for RIF), which is CYP3A4 dependent. The other hydroxylated metabolites (16alpha-OH and 2alpha-OH) were also enhanced. The metabolism of MDZ, which is specifically metabolized by CYP3A4 in humans, was also investigated following drug's exposure to hepatocytes. DZP one, which is governed by various CYPs, including CYP3A, was also investigated. RBT was shown to increase the biotransformation of both benzodiazepines (approximately 1.9-fold over control). Moreover, the effects of both drugs on ethoxyresorufin O-deethylase activity (EROD), which is representative of CYPIA1/2 isoforms, were tested. Results showed only a moderate induction of this marker (approximately 2-fold over control) when compared to the high effect observed after hepatocyte exposure to 3-methylcholantene (approximately 14-fold over control). Finally, the action of RBT and RIF on UGTs expression was investigated by using AZT as diagnostic substrate: glucuronides formation was not significantly affected by the two rifamycin derivatives. On the whole, exposure of fresh or cryopreserved human hepatocytes to RBT dose-dependently affected the levels of drug metabolizing enzymes in a dose-dependent manner. However, as already demonstrated by in vivo pharmacokinetic studies, its inducing properties towards CYPs, CYP3A in particular, are less pronounced than RIF.

Publication types

  • Comparative Study

MeSH terms

  • Aryl Hydrocarbon Hydroxylases*
  • Cryopreservation*
  • Cytochrome P-450 CYP1A1 / biosynthesis
  • Cytochrome P-450 CYP3A
  • Cytochrome P-450 Enzyme System / biosynthesis*
  • Diazepam / metabolism
  • Diazepam / pharmacology
  • Dose-Response Relationship, Drug
  • Enzyme Induction / drug effects
  • Glucuronosyltransferase / biosynthesis*
  • Humans
  • Liver / drug effects*
  • Liver / enzymology
  • Midazolam / metabolism
  • Midazolam / pharmacology
  • Organ Preservation*
  • Oxazines / metabolism
  • Oxazines / pharmacology
  • Oxidoreductases, N-Demethylating / biosynthesis
  • Rifabutin / pharmacology*
  • Rifampin / pharmacology*
  • Testosterone / metabolism
  • Testosterone / pharmacology
  • Zidovudine / metabolism
  • Zidovudine / pharmacology


  • Oxazines
  • Rifabutin
  • Testosterone
  • Zidovudine
  • ethoxyresorufin
  • Cytochrome P-450 Enzyme System
  • Aryl Hydrocarbon Hydroxylases
  • Cytochrome P-450 CYP1A1
  • Cytochrome P-450 CYP3A
  • Oxidoreductases, N-Demethylating
  • Glucuronosyltransferase
  • Diazepam
  • Midazolam
  • Rifampin