In this study, enzyme-deep eutectic solvent-assisted ultrasonic extraction technique (EnDUE) was developed for the efficient dissolution of flavonoids from Artemisiae Argyi Folium. The extraction results of Artemisiae Argyi Folium flavonoids (quercetin, luteolin, and isorhamnetin) were used as indicators to investigate the influencing factors through single factor experiment, Placket-burman design, and Box-behnken design, so as to obtain satisfactory yields. After systematic optimization, the optimal conditions for extraction of the target flavonoids were: Choline chloride/1,4-butanediol with a water content of 25%, cellulase+pectinase with a concentration of 1.6%, solid-liquid ratio of 1/32 g/mL, pH of 4.2, ultrasonic frequency of 80 kHz, ultrasonic power of 160 W, ultrasonic temperature of 40 °C, and ultrasonic time of 25 min, respectively, which derived a total yield of 8.06 ± 0.29 mg/g. Compared with the reference techniques, the proposed EnDUE technique showed significant advantages in the yield and extraction efficiency of flavonoids. In addition, after preliminary purification, the Artemisiae Argyi Folium flavonoids showed good antioxidant activity. Deep eutectic solvent (DES) can degrade the cell wall components and increase the action site of enzyme, and enzyme can promote the penetration of DES into the cell wall matrix, which is mutually beneficial to the dissolution of intracellular components. Therefore, the extraction technique proposed in this work (EnDUE) greatly promotes the dissolution of flavonoids from Artemisiae Argyi Folium, and provides theoretical support for the further application of plant flavonoids.