Non-alcoholic fatty liver disease (NAFLD) is an increasingly recognized condition that encompasses a spectrum of liver abnormalities. It has been suggested that oxidative stress and lipid peroxidation are key pathophysiological mechanisms in NAFLD. Although an antioxidant effect of apolipoprotein A-I (apoA-I) has been reported, its influence on NAFLD has not been reported. The aim of this study was to determine whether apoA-I could improve the biochemical and histological abnormalities associated with high-fat diet-induced NAFLD through its antioxidant actions in rabbits. Liver damage was evaluated by hepatic coefficient, hepatic lipid assay, liver apparent abnormalities as well as hematoxylin-eosin staining of liver sections. Lipid peroxidation was assessed by measuring malondialdehyde (MDA) level in liver. Oxidative stress was assessed by measuring superoxide dismutase (SOD), glutathione peroxidase (GPx), and inducible nitric oxide synthase (iNOS) activities in serum and liver. Also, the mRNA expressions levels of SOD, GPx, and catalase (CAT) were determined by real-time quantitative polymerase chain reaction method. The results showed that apoA-I (20 or 40 mg/kg/w) was effective in reducing hepatic steatosis, inflammation, hepatic coefficient, and liver total cholesterol, triglyceride, low-density lipoprotein-cholesterol, and MDA levels in high-fat diet rabbits. In addition, apoA-I increased SOD and GPx activities while reducing iNOS activity in serum and liver. Moreover, apoA-I significantly increased the mRNA expression levels of SOD, GPx, and CAT in liver. This study showed that apoA-I exerted protective effects against fatty liver disease in rabbits induced by a high-fat diet, possibly through its antioxidant actions.