The aim of the present study was to investigate the effects of fermented barley supplementation on blood lipid profiles, carnitine concentrations, and hepatic mRNA levels of enzymes involved in fatty acid metabolism. Thirty-two C57BL/6J male mice were divided into 4 groups; normal diet control group (ND), high fat diet control group (HD), high fat diet plus barley supplemented group (BR), and high fat diet plus fermented barley supplemented group (BR-F). BR-F supplementations decreased total cholesterol (TC) and LDL-cholesterol in serum, triglyceride (TG) in liver. Serum total carnitine (TCNE) concentrations were significantly higher in the BR-F group than HD group. BR-F supplementations significantly increased hepatic lipolysis regulating gene expression such as carnitine palmitoyltransferase-I (CPT-I), peroxisome proliferator-activated receptor-${alpha}$ ($PPAR{alpha}$), whereas significantly decreased lipogenic enzyme expression such as acetyl CoA carboxylase (ACC). These results suggest that the fermented barley has anti-obesity properties.