Xenobiotic metabolism by cultured primary porcine hepatocytes.

Publication Type:

Journal Article

Source:

Tissue Eng, Volume 6, Issue 5, p.467-79 (2000)

Keywords:

7-Alkoxycoumarin O-Dealkylase, Albumins, Animals, Cell Separation, Cells, Cultured, Coumarins, Cytochrome P-450 CYP1A1, Cytochrome P-450 CYP2B1, Cytochrome P-450 Enzyme System, Diazepam, Dicumarol, Dose-Response Relationship, Drug, Enzyme Induction, Enzyme Inhibitors, Glucuronosyltransferase, Hepatocytes, Humans, Inactivation, Metabolic, Liver, Oxazines, Oxidoreductases, Phenolsulfonphthalein, Rats, Rats, Inbred Lew, Swine, Urea, Xenobiotics

Abstract:

<p>Considering the large yield of viable cells comparable to human liver, primary porcine hepatocytes offer a valuable resource for constructing a bioartificial liver device. In this study, the ability of cultured primary porcine hepatocytes to detoxify xenobiotics has been examined using various known substrates of cytochrome P450 isoenzymes and UDP-glucuronosyltransferases. Present investigation demonstrated the stability of the isoenzymes responsible for the metabolism of diazepam in native state and stabilization of other isoenzymes, as judged by ethoxycoumarin o-dealkylase (ECOD), ethoxyresorufin o-dealkylase (EROD), benzyloxyresorufin o-dealkylase (BROD), and pentoxyresorufin o-dealkylase (PROD) activities following induction in culture environment, for a period of 8 days. Resorufin O-dealkylase activities were found to be the most unstable and deteriorated within first 5 days in culture. These activities were restored following induction with 3-methylcholanthrene (3-MC) or sodium phenobarbital (PB) to 20-fold of 1 activity for EROD, and 60 and 174% of day 1 activity for PROD and BROD on day 8, respectively. Metabolism of methoxyresorufin was most strikingly increased following induction with 3-MC to approximately 60-fold of day 1 activity, on day 8. UDP-glucuronosyltransferase-dependent glucuronidation of phenol red, however, stayed intact during the course of our study without induction. Our study indicated that porcine hepatocytes in vitro maintain many important liver-specific functions including detoxification (steady state and inducibility).</p>

PDF: