Non-alcoholic hepatic steatosis is a phenotype of metabolic syndrome, and aging is a risk factor for this condition. Senescence-accelerated mouse prone 8 (SAMP8) is a murine model for studying aging-associated disorders. We here investigated the effect of dietary supplementation with L -lysine (Lys) on non-alcoholic hepatic steatosis in SAMP8 mice. Triglyceride (TG) and cholesterol (Chol) accumulated in the livers of SAMP8 mice fed a standard diet at 36 wk of age. However, intake of a Lys-rich diet for 2 mo prevented the accumulation of TG and Chol in the liver. Plasma alanine aminotransferase activity, an index of liver injury, was decreased by Lys. The mRNA expression levels of peroxisome proliferator-activated receptor gamma coactivator 1-α and carnitine palmitoyltransferase 1a, which regulate β-oxidation, were increased in the livers of SAMP8 mice fed the Lys-rich diet. Taken together, our study suggests dietary intake of Lys prevents hepatic steatosis by stimulating β-oxidation in SAMP8 mice.