Perfluorododecanoic acid (PFDoA), a perfluorinated carboxylic acid (PFCA) with twelve carbon atoms, has broad industrial applications and is widely distributed in both wildlife and the environment. Unlike other PFCAs with short carbon chain, however, limited studies have been performed to date on the toxic effects of PFDoA on animals. To determine the hepatotoxicity of PFDoA, male rats were orally dosed by gavage for 14 days with 0, 1, 5, or 10 mg PFDoA/kg/day. Absolute liver weights were diminished, but the relative liver weight was significantly increased in the 5 and 10 mg PFDoA/kg/day groups. Meanwhile, serum triglyceride (TG) concentrations were decreased significantly in rats dosed with 1 and 5 mg PFDoA/kg/day, while the liver lipid accumulation was observed in ultrastructure. The expression of peroxisome proliferator-activated receptor (PPAR)-alpha and its target genes, and to a lesser extent PPARgamma, was induced by PFDoA. No significant changes in the expression of liver X receptor alpha (LXRalpha) or its target genes CYP7A1 and acetyl-CoA carboxylase 1 (ACC1) were noted, although the mRNA levels of several genes involved in lipogenesis and lipid transport were changed significantly in the certain of the experimental groups. In addition, superoxide dismutase (SOD) and catalase (CAT) activities were activated significantly in the 1 mg PFDoA/kg/day group and inhibited significantly with a concomitant increase of lipid peroxidation (LPO) levels in the 5 and 10 mg PFDoA/kg/day groups. Our results demonstrate that PFDoA exerts notable hepatotoxicity in male rats and that PPAR and its target genes, SOD and CAT activity, and LPO levels exhibited sensitivity to the toxicity of PFDoA.