Differential Response of Cultured Human Umbilical Vein and Artery Endothelial Cells to Ah Receptor Agonist Treatment: CYP-Dependent Activation of Food and Environmental Mutagens

Abstract

In the present study, 7-ethoxyresorufin O-deethylase (EROD), 7,12-dimethylbenz[a]anthracene (DMBA)-hydroxylase, and covalent binding of 3H-labeled 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (3H-Trp-P-1) and 3H-DMBA were examined in human umbilical vein endothelial cells (HUVEC) and human umbilical artery endothelial cells (HUAEC) exposed to the aryl hydrocarbon (Ah) receptor agonist β-naphthoflavone (BNF) or vehicle only. The results revealed a marked induction of enzymatic activity in BNF-treated HUVEC compared with vehicle-treated cells, whereas no similar response was observed in BNF-treated HUAEC. EROD, DMBA hydroxylase, and covalent binding of 3H-Trp-P-1 and 3H-DMBA in BNF-treated HUVEC were reduced in the presence of the CYP1A inhibitor ellipticine. Addition of other CYP1A inhibitors α-naphthoflavone, miconazole, 1-ethynylpyrene, 1-(1-propynyl)pyrene), or the CYP1A substrate ethoxyresorufin to the incubation buffer of BNF-treated HUVEC reduced covalent binding of 3H-Trp-P-1 by 93–98%. Western blot analysis confirmed an induction of CYP1A1 in BNF-treated HUVEC, but not in BNF-treated HUAEC. CYP1A1 was, however, detected in both vehicle- and BNF-treated HUAEC. The results showed that BNF exposure induced CYP1A1 and metabolic activation of xenobiotics in HUVEC, whereas the catalytic activity remained low in BNF-treated HUAEC. Our results suggest that endothelial lining of human veins may be a target for adverse effects of xenobiotics activated into reactive metabolites by Ah receptor-regulated enzymes. Several studies have detected CYP1A1 in endothelial linings, whereas expression of CYP1A2 and CYP1B1 seems to be negligible at this site. This suggests that the metabolic activation and covalent binding of 3H-Trp-P-1 and 3H-DMBA in HUVEC are most likely mediated by CYP1A1.