Kupffer cell-mediated differential down-regulation of cytochrome P450 metabolism in rat hepatocytes

Eur J Pharmacol. 1999 Feb 26;368(1):75-87. doi: 10.1016/s0014-2999(98)00988-1.


Nonparenchymal cells, particularly Kupffer cells, might play an important role in the modulation of xenobiotic metabolism in liver and its pharmacological and toxicological consequences. This intercellular communication via the exchange of soluble factors was investigated in primary rat Kupffer cells and hepatocytes. Freshly isolated rat Kupffer cells were seeded onto cell culture inserts and cocultured with 5 day old serum-free rat hepatocyte monolayer cultures at a ratio of 1:1 for 2 days. Hepatocyte cultures, Kupffer cell cultures or cocultures were treated with 0.1 ng/ml-10 microg/ml lipopolysaccharide (LPS). Within this concentration range, no significant toxicity was observed in either cell type. In LPS-exposed cocultures, tumor necrosis factor alpha (TNFalpha) levels rose up to 5 ng/ml within 5 h; nitric oxide (NO) levels increased up to 70 microM within 48 h of treatment, both in a dose-dependent fashion. The release of negative (albumin) and positive (alpha1-acid-glycoprotein) acute phase proteins from the hepatocytes was strongly down- and up-regulated, respectively. The simultaneous treatment of the cocultures with phenobarbital and LPS (10 ng/ml) or 3-methylcholanthrene and LPS (10 ng/ml) resulted in a strong down-regulation (85%) of the phenobarbital-induced cytochrome P450 (CYP) isoform CYP2B1 in the hepatocytes whereas the 3-methylcholanthrene-induced isoform CYP1A1 was only weakly affected (15%). This specific down-regulation of CYP2B1 was mediated exclusively by TNFalpha, released from the Kupffer cells. It was not linked with NO release from or inducible NO synthase activity in the hepatocytes. The TNFalpha release was not affected by the two xenobiotics. Acetaminophen tested in these cocultures showed no direct interaction with the Kupffer cells. The use of liver cell cocultures is therefore a useful approach to investigate the influence of intercellular communication on xenobiotic metabolism in liver.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetaminophen / pharmacology
  • Acute-Phase Proteins / drug effects
  • Acute-Phase Proteins / metabolism
  • Albumins / drug effects
  • Albumins / metabolism
  • Analgesics, Non-Narcotic / pharmacology
  • Animals
  • Carcinogens / pharmacology
  • Cell Survival / drug effects
  • Cells, Cultured
  • Coculture Techniques
  • Cytochrome P-450 Enzyme System / drug effects
  • Cytochrome P-450 Enzyme System / metabolism*
  • Dose-Response Relationship, Drug
  • Down-Regulation
  • Isoenzymes / drug effects
  • Isoenzymes / metabolism
  • Kupffer Cells / cytology
  • Kupffer Cells / drug effects
  • Kupffer Cells / physiology*
  • Lipopolysaccharides / pharmacology
  • Liver / cytology
  • Liver / drug effects
  • Liver / enzymology*
  • Male
  • Methylcholanthrene / pharmacology
  • Nitric Oxide / metabolism
  • Phenobarbital / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Tumor Necrosis Factor-alpha / drug effects
  • Tumor Necrosis Factor-alpha / metabolism
  • Xenobiotics / pharmacology


  • Acute-Phase Proteins
  • Albumins
  • Analgesics, Non-Narcotic
  • Carcinogens
  • Isoenzymes
  • Lipopolysaccharides
  • Tumor Necrosis Factor-alpha
  • Xenobiotics
  • Nitric Oxide
  • Acetaminophen
  • Methylcholanthrene
  • Cytochrome P-450 Enzyme System
  • Phenobarbital