Physiological properties of homogenized and non-homogenized polysaccharide from the seeds of Plantago asiatica L., including antioxidant capacity and short-chain fatty acid (SCFA) production, were compared in this study. High pressure homogenization decreased particle size of the polysaccharide, and changed the surface topography from large flake-like structure to smaller porous chips. FT-IR showed that high pressure homogenization did not alter the primary structure of the polysaccharide. However, high pressure homogenization increased antioxidant capacity of the polysaccharide, evaluated by 4 antioxidant capacity assays (hydroxyl radical-scavenging, superoxide radical-scavenging, 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH)-scavenging and lipid peroxidation inhibition). Additionally, the production of total SCFA, propionic acid and n-butyric acid in ceca and colons of mice significantly increased after dieting supplementation with homogenized polysaccharide. These results showed that high pressure homogenization treatment could be a promising approach for the production of value-added polysaccharides in the food industry.