The Zanthoxylum bungeanum leaf has been traditionally taken as a vegetable and seasoning in China.To disclose the mechanisms of its benefit for human health, antioxidant activity and cytoprotective effect of the leaf were investigated. The ethyl acetate fraction exhibited the strongest ABTS and DPPH radicals scavenging activities as compared with chloroform and water fractions, as well as the crude extract. This fraction exhibited a protective effect against hydrogen peroxide-induced cytotoxicity in PC12 cells by MTT reduction assay. Furthermore, the cytoprotective effect was confirmed using flow cytometry analysis. Comparing with the model cells, the ethyl acetate fraction significantly increased G0/G1 and G2/M phase cells together with reducing S phase cells. Ten flavonoid glycosides were identified as isovitexin, vitexin, hyperoside, rutin, isoquercitrin, foeniculin, trifolin, quercitrin, astragalin, and afzelin from this fraction using high performance liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance spectroscopy. This is the first report for the presence of vitexin, isovitexin, astragalin, trifolin and afzelin in Z. bungeanum. Hence, the results present here suggest the potential utility of Z. bungeanum leaf as a source of natural antioxidant due to its diversity and high content of flavonoids.