In this study, high-fat diet-induced obese mice were administered oat soluble (SOG) and insoluble β-glucan (IOG) at different doses. Mice were sacrificed after 6 weeks, body weight, serum lipid level, fecal pH value, fecal bile acid excretion and total colonic short-chain fatty acid (SCFA) concentration was measured. Mesenteric adipocyte count and size were also evaluated histologically. The population of Lactobacillus in colon was determined. Compared with obese mice administered normal saline, body weight, serum cholesterol, triglycerides and the lipoprotein profile were significantly decreased (p<0.05) in mice administered SOG and IOG. The fecal pH value was significantly decreased (p<0.05), while both fecal bile acid excretion and total colonic SCFA concentration were significantly increased (p<0.05). In addition, it exhibited an increase of fat cell count and a decrease of cell size. SOG and IOG restored the number of Lactobacillus in colon (p<0.05). IOG was more effective on weight-loss while SOG might play a more important role in improving serum lipids and the efficacy of promoting growth of Lactobacillus is similar. A dosage of 2 g⋅kg -1 ⋅BW (Body Weight) produced the most significant effect. These data were anticipated to support the prebiotic property and anti-obesity effect of oat β-glucan.