The effects of different extraction methods on extraction rate, structure, antibacterial activity and antioxidant activity of horse bone marrow protein were studied, and its edible and medicinal values were examined. Twelve extraction methods were screened through using protein content, extraction rate, molecular weight and antioxidant activity as the indicators, and structural characterizations were performed by UV, SDS-PAGE, FT-IR, and SEM-EDX analysis. The biological properties of the obtained proteins were evaluated by in vitro antibacterial and antioxidant activities. After comprehensive investigations, it was found that the index scores of the horse bone marrow proteins extracted with water, 0.25% NaCl, and normal saline were higher, with their corresponding protein contents being 41.91, 39.93 and 37.82 mg/mL, and extracted rates being 19.26%, 14.86% and 23.22%, respectively. UV spectra showed that the horse bone marrow proteins prepared by different methods had the maximum absorption peak around 270 nm. SDS-PAGE revealed that the molecular weights of the obtained proteins were mainly around 66 ku, with the presence of corresponding polypeptide bands. FT-IR spectra showed typical protein absorption signals in the region of 4000~400 cm-1. SEM-EDX analysis showed that the obtained proteins were composed of spherical, striped and irregular particles. The results of biological activity experiments demonstrated that the horse bone marrow proteins prepared by the three methods exhibited sound antibacterial and antioxidant effects, and the zone diameters of inhibition against Staphylococcus aureus were 12.0, 16.0 and 11.0 mm, respectively. The IC50 values of DPPH and ABTS free radicals scavenging activities were 0.32, 0.29 and 0.31 mg/mL, and 0.26, 0.15 and 0.20 mg/mL, respectively. The research results can provide a scientific basis for further development and utilization of horse meat processing by-products, bone resources. Key words: