Cytochrome P4501A1 mediates the metabolism of 2,3,7,8-tetrachlorodibenzofuran in the rat and human

Previous studies have established that TCDF is rapidly metabolized and excreted in rats and that pretreatment of rats with TCDD increases the rate of hepatic metabolism of this compound. The extrahepatic metabolism of TCDF was investigated to assess which enzyme was involved in the metabolism of this compound. Very little metabolism of TCDF was detected in control microsomes (0.3-3.0 pmol/mg/hr), while TCDF metabolism was increased 40- to 200-fold in TCDD-induced rat liver, kidney, and lung microsomes. Since TCDD induces cytochrome P4501A1 and P4501A2 (CYP1A1 and CYP1A2) in the rat liver but only CYP1A1 in kidney and lung, these results suggest that CYP1A1 metabolizes TCDF. To test this hypothesis, TCDF metabolism was investigated in the presence and absence of selective chemical inhibitors and antibodies to CYP1A1 and 1A2. 1-Ethynylpyrene, a suicide inhibitor of CYP1A1 and antibody to rat CYP1A1, produced a dose-dependent inhibition of TCDF metabolism in TCDD-induced rat liver microsomes. Conversely, 2-ethynylnaphthalene, a suicide inhibitor of CYP1A2 and antibody to rat CYP1A2, had no inhibitory effect on the hepatic microsomal metabolism of TCDF. Together, the results strongly indicate that rat CYP1A1 is the primary enzyme responsible for the metabolism of TCDF. 4-Hydroxy-2,3,7,8-TCDF was also identified as the major TCDF metabolite formed by rat CYP1A1. TCDF was also metabolized by human liver microsomes and recombinant yeast microsomes expressing human CYP1A1 and reductase but not by yeast microsomes expressing human CYP1A2 with or without reductase. A similar HPLC profile of TCDF metabolites was observed with microsomes from human liver and yeast expressing human CYP1A1. However, based on ethoxyresorufin-O-deethylase activity, a marker of CYP1A1, the relative rate of TCDF metabolism is about 100-fold greater in TCDD-induced rat liver microsomes than in yeast microsomes expressing human CYP1A1 and reductase. Thus, although TCDF is metabolized by rat and human CYP1A1, the results indicate that there are marked quantitative differences in metabolism which suggest that TCDF will be more persistent in humans