The results suggested that induction of peroxisomal β‐oxidation plays a critical role in ethanol‐induced hepatic TG accumulation in animals under a fasted state, while specific inhibition of ACOX‐1, the first enzyme in peroxisomal β‐oxidation that generates hydrogen peroxide, improved alcohol‐induced hepatic steatosis in the fasting mice. This evidence concerns the gene ACOX1 and fatty liver disease.