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The present study examined the anti-obesity effects of Pinus sylvestris bark (PS). Anti-obesity activity of PS (286 and 667 mg/kg/day for 56 days) evaluated using High fat diet induced obesity (58.23% of energy as Fat) in C57BL/6J female mice. We also investigated the effect of PS extract on adipocyte differentiation of 3T3-L1 preadipocytes and In-vitro pancreatic lipase inhibitory activity. Oral treatment with PS ethanol extracts for a period of 56 days, resulted in significant precluded the increase in body weight, adipose tissue weight and serum triglycerides, total cholesterol, leptin and insulin levels in mice with obesity induced by a high-fat diet. PS extract showed the highest and significant inhibitory effect (28.00±0.86%) at concentration of 250 µg/ml on pancreatic lipase activity. The results of in vitro cytotoxicity assay also showed that PS efficiently inhibited the proliferation in 3T3-L1 preadipocytes. In conclusion, Pinus sylvestris bark can be used for an obesity preventing or therapeutic agent because the extract can suppress the adipocyte differentiation, decrease body weight and body fat in an obese animal and inhibition of intestinal lipid absorption thereby suppressing fat accumulation.