Abstract: In this study, Laoxianghuang polysaccharide was used as the experimental material to examine the protective effects on stomach mucosa. The polysaccharide was first extracted by hot water and then purified by Sevag methods deproteinization, DEAE-52 cellulose anion-exchange chromatography column, and CL-6B agarose gel column. The ethanol-damaged human gastric mucosal epithelial cells (GES-1) model and the GES-1 scratch injury model were used to evaluate gastric mucosal protective activity of the polysaccharide in vitro. Results showed that six fractions of Laoxianghuang polysaccharide were obtained, including PFCP, PFCP-1, PFCP-2, PFCP-3, PFCP-2-1, and PFCP-2-2. After ethanol damaged to GES-1, each fraction of Laoxianghuang polysaccharide improved cellular activity, increased the activity of superoxide dismutase (SOD), catalase (CAT), and inhibited malondialdehyde (MDA) content, exhibiting good antioxidant capacity to reduce mucosal damage. Among them, PFCP-2-1 (50 μg/mL) had the best effect on improving SOD enzyme activity and reducing MDA content, and PFCP-2-1 (5 μg/mL) had the best effect on improving CAT enzyme activity, which was significant compared with the model group (P<0.05) and better than the teprenone-positive drug group. It was demonstrated that Laoxianghuang polysaccharide all fractions accelerated gastric epithelial cell growth, promoted cell migration to the damaged area, and restored gastric mucosal epithelial cell integrity in the GES-1 scratch injury assay. Positive drugs had the most effective scratch repair rate, followed by PFCP-2-1（50 μg/mL）, but there was no significant difference between them (P>0.05). In conclusion, Laoxianghuang polysaccharide PFCP-2-1 may exert gastric mucosal protective activity by enhancing cellular antioxidant capacity as well as promoting the proliferation and migration of GES-1.