Objective To explore the preparation conditions, structural information, and in vitro hypoglycemic activities of acetylated Elaeagnus moorcroftii Wall. polysaccharides of Kashgar (A-EMP-K).Methods EMP-K is extracted by the water extraction and alcohol precipitation method, and modified by the acetic anhydride method. Then, A-EMP-K preparation conditions are optimized using Box-Behnken response surface methodology. Furthermore, structural characterization and in vitro hypoglycemic activity assays of A-EMP-K are performed.Results The optimal preparation conditions are a reaction time of 3 h, a reaction temperature of 61 ℃, and an acetic anhydride addition volume of 9.3 mL. Under these conditions, A-EMP-K exhibits a degree of substitution at 0.326. The A-EMP-K is a heteropolysaccharide consisting of galactose, arabinose, galacturonic acid, and other constituents. The absorption peaks of infrared spectra at 1 731 cm^-1 and 1 257 cm^-1 confirm the successful introduction of acetyl groups. Scanning electron microscope (SEM) results reveal a distinct porous structure and a relatively smooth surface. The particle size and zeta potential results indicate a relatively stable solvent system. With acarbose as the positive control, the inhibition rate of the A-EMP-K is (63.80±4.58)% against α-glucosidase, and is (81.18±2.48)% against α-amylase.Conclusion EMP-K and A-EMP-K exhibit potent performance in hypoglycemic activities.