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This study investigated the role of ethanol‐inducible cytochrome P450‐2E1 (CYP2E1) in enhancing CYP2E1 and other P450 proteins in extracellular vesicles (EVs) from alcohol‐exposed rodents and human patients with alcoholism and their effects on oxidative hepatocyte injury. Female Fischer rats and wild‐type or Cyp2e1‐null mice were exposed to three oral doses of binge ethanol or dextrose control at 12‐hour intervals. Plasma EV and hepatic proteins from alcohol‐exposed rodents, patients with alcoholism, and their respective controls were isolated and characterized. The number of EVs and the amounts of EV CYP2E1, CYP2A, CYP1A1/2, and CYP4B proteins were markedly elevated in both patients with alcoholism and alcohol‐exposed rats and mice. The number of EVs and EV P450 proteins were significantly reduced in ethanol‐exposed rats fed a diet containing polyunsaturated fatty acids. The …