The invention relates to a method of extracting peony seed oil from peony seeds. The method includes following process steps: (1), crushing and sieving shelled peony seeds to obtain peony seed powderparticles smaller than 40 meshes in particle size; (2), taking peony seed powder, adding water, and mixing well to obtain mixed liquid; (3), adjusting pH value of the mixed liquid, stirring at certaintemperature for a period of time during water bath, and performing acid heat process wall breaking; (4), adding medium-temperature amylase into obtained mixed liquid, and performing enzyme deactivation after enzymolysis; (5), adding neural protease into the mixed liquid after enzyme deactivation, performing enzyme deactivation after enzymolysis, and centrifugally separating to obtain free oil, anemulsified layer, hydrolysate and residue; (6), taking the emulsified layer for freeze-demulsifying, unfreezing, centrifugally separating to obtain an oil phase, and combining the oil phase with thefree oil obtained in the step (5) to obtain the peony seed oil. Acid process wall breaking is combined with joint action of medium-temperature amylase and neural protease, so that total oil extractionrate can reach 86.6%.