PineXol, extracted from Korean red pine bark, has beneficial effects, such as antioxidant, antiinflammatory, and antilipogenic activities in vitro. We tested the hypothesis that PineXol supplementation could have anti-obesity effects on mice fed a high-fat diet (HFD). Four-week-old male C57BL/6 mice were fed normal chow (18% kcal from fat) or a HFD (60% kcal from fat). HFD-fed animals were also subjected to PineXol treatment at a dose of 10 or 50 mg/kg body weight (BW) (PX10 or PX50, respectively) body weight. The body weight and body fat mass in the PX50 group were statistically lower than those in the HFD group ( < 0.05 and < 0.001, respectively). The concentration of hepatic triglycerides, total cholesterol, and low-density lipoprotein cholesterol were reduced in the PX50 group compared with the HFD group ( < 0.01). Acetyl CoA carboxylase ( < 0.01), elongase of very long chain fatty acids 6 ( < 0.01), stearoyl CoA desaturase 1 ( < 0.05), microsomal triglyceride transfer protein ( < 0.01), and sterol regulatory element-binding protein 1 ( < 0.05) were significantly decreased in the PX50 group compared with that in the HFD group. In white adipose tissue, CCAATenhancer-binding protein alpha ( < 0.05), peroxisome proliferator-activated receptor gamma ( < 0.001), and perilipin ( < 0.01) were decreased in the PX50 group compared with those in the HFD group. Therefore, the current study implies the potential of PineXol for the prevention and/or amelioration of obesity, in part by inhibition of both hepatic lipid synthesis and adipogenesis in white adipose tissue.