Female wild type SV129 mice, CYP2E1 knockout mice (KO) and CYP2E1 knockin mice (KI) in which the human CYP2E1 was added to replace the knocked out mouse CYP2E1 were fed ethanol for three weeks. Pair-fed controls received isocaloric dextrose. All mice were killed after an overnight fast. Serum ethanol levels were similar between the 3 groups of ethanol-fed mice at sacrifice. Ethanol elevated mouse CYP2E1 levels (Fig 1A) and CYP2E1 catalytic activity (oxidation of p-nitrophenol) (Fig 1B) about two-fold in wild type mice. Levels of CYP2E1 and oxidation of PNP were either very low or not detectable in the dextrose or ethanol-fed KO mice (Fig. 1A, 1B). Levels and activity of human CYP2E1 were elevated in the ethanol-fed KI mice to an even greater extent than that found for mouse CYP2E1 in the ethanol-fed WT mice (Fig. 1A,1B). Immunohistochemistry showed that chronic ethanol feeding elevated CYP2E1 in WT mice and more strikingly in KI mice in the pericentral zone of the liver acinus. No CYP2E1 was observed in the ethanol- or dextrose-fed KO mice (Fig. 1C).. Hepatic levels of TNFα were not significantly different between the ethanol and dextrose fed mice and between the three genotypes (Fig. 1D).
