THE EFFECT OF ETHANOL ON COUMARIN METABOLISM AND CYTOTOXICITY IN TRANSFECTED CULTURES OF HEPG2 CELLS WITH CYP2E1 EXPRESSION.

Jabłoński, Jakub; Jabłońska, Ewa; Hołownia, Adam; Moniuszko-Jakoniuk, Janina

Introduction: Coumarin is present in a number of food products. Due to the fact that this compound is used for aromatising alcoholic beverages and, with the participation of ethanol-induced P-450 CYP2E1 isoform, may be metabolised to potentially toxic metabolites, it was decided to determine the extent to which ethyl alcohol affects coumarin metabolism in the transfected HepG2 hepatoblastoma cells with CYP2E1 expression. Material and Methods: The study was performed on HepG2 cell cultures with CYP2E1 expression (E47). Cells transfected with an empty vector (C34) were used as control. Cultures were maintained at coumarin concentrations of 0.1 mg/ml and 1 mg/ml, with 100 mM concentration of coumarin and ethanol. The denomination of coumarin concentration in the culture supernatants were made with use of gas chromatography. Cell vivacity and apoptosis were estimated by means of flow cytometry. Results: It was established that, compared to C34 cells, E47 cells exhibit a higher metabolic activity towards coumarin, with ethanol increasing this activity. In E47 cell cultures with coumarin at a concentration of 0.1 mg/cm3 and ethanol, an increase in the percentage of apoptotic cells was shown. In cell cultures with coumarin at a concentration of 1 mg/ml, a substantial effect of coumarin on the increased percentage of dead and apoptotic cells was evidenced. This concerned both C34 and E47 cell cultures. The changes described here were more intensive in E47 cell cultures. Conclusions: In conclusion it can be assumed that ethanol increases coumarin metabolism to toxic metabolites by inducing the expression of CYP2E1 isoenzyme.

COUMARINS; CELL-mediated cytotoxicity; CYTOMETRY; ALCOHOLIC beverages; CHROMATOGRAPHIC analysis; APOPTOSIS; ISOENZYMES

Acta Toxicologica, 2007, Vol 15, Issue 2, p117

1731-6383

Academic Journal