Abstract: Total flavonoids of Hanyuan Zanthoxylum bungeanum were prepared by ultrasonic-assisted solvent extraction and macroporous resins purification in this study, and the anti-oxidative activity in vitro as well as anti-proliferative activity on Hela cells was studied. The anti-oxidative activity in vitro was detected by the scavenging rates of DPPH·and hydroxyl radicals, the formation of phosphorus molybdenum complex and the reducing ability of Fe³⁺. The anti-proliferative activities on Hela cells were detected by CCK-8 as well as the fluorescent staining methods of PI and EGFP-Annexin V/PI. The results showed that the yield of total flavonoids was 2.19%. The scavenging rates of DPPH and hydroxyl radicals were 84.70% and 83.61% respectively at the concentrations of 125 and 500 μg/mL, and the IC₅₀ values were 65.61 and 194.40 μg/mL respectively. The reducing power and total anti-oxidative capacity were the strongest at the concentrations of 400 and 300 μg/mL, and the absorbance of reducing power and total antioxidant capacity were 0.70 and 0.48, respectively. In the study of cells, the viabilities of Hela cells were 27.15%, 10.43% and 30.63% respectively after 12, 24, and 48 h treated by the concentration of 800 μg/mL, and decreased significantly (P<0.001) compared to the control at the same time, which could be due to cell cycle arrest induced in G₂/M phase and the cell apoptosis rate increased at this mass concentration at 24 h.Therefore, the total flavonoids extract from Hanyuan Zanthoxylum bungeanum had the anti-oxidative activity in vitro and anti-proliferative activity on Hela cells, which could be theoretical support for further using Hanyuan Zanthoxylum bungeanum as a source of some functional products.