Abstract: OBJECTIVE To explore the antioxidant molecular mechanism of phenylethanoid glycosides from Cistanches Herba by employing the network pharmacological method, as well as to explore the antioxidant activity and structure-activity relationship of the main phenylethanoid glycosides by in vitro experiments. METHODS Phenylethanoid glycosides from 5 Cistanches Herba distributed in China were obtained by TCMSP database and literature search. Pharmmapper web server, CoolGeN and GeneCards database were used to predict and screen the antioxidant targets of the above components and the ingredient-target network was subsequently constructed by Cytoscape 3.6.1. The protein-protein interaction network was constructed by the String database. The GO and KEGG pathway analysis of the targets were analyzed by the DAVID database. The ingredient-target-pathway-disease network was constructed by Cytoscape 3.6.1. DPPH radical scavenging method and Prussian blue spectrophotometry were used to evaluate the antioxidant capacity and total reducing capacity of phenylethanoid glycosides, aglycons and structural analogs, the molecular docking was adopted to evaluate the binding ability of the above compounds and important targets, providing a basis for target prediction and validation of the ingredients. RESULTS It was found that 71 phenylethanoid glycosides active components and 114 targets were involved. A total of 44 antioxidant-related targets were further screened, including glutathione S-transferase P1(GSTP1), epidermal growth factor receptor(EGFR). KEGG pathway analysis showed that the above antioxidant-related targets were enriched in 51 pathways, such as the prolactin signaling pathway, FoxO signaling pathway, and Fc epsilon RI signaling pathway. Four phenylethanoid glycosides, aglycons and structural analogs had the ability to scavenge DPPH free radicals and strong reducing power, also had strong binding ability with the targets. CONCLUSION Phenylethanoid glycosides can exert antioxidant effects by modulating GSTP1, EGFR, MAPK1 and other targets. The antioxidant activities of phenylethanoid glycosides are related to caffeoyl and 3,4-dihydroxyphenylethanol groups in the structure.