Research interest in examining Elaeagnus angustifolia's potential as a source of anti‐inflammatory and antioxidant agents has grown as a result of the plant's endorsement as a rich source of bioactive chemicals with promising anti‐inflammatory and antioxidant activity. In this study, zinc oxide (Fe0.25‐ZnO) bimetallic nanoparticles (E.ang‐Fe0.25‐ZnO NPs) were synthesized using an aqueous extract of Elaeagnus angustifolia. Synthesized Fe0.25‐ZnO nanoparticles were characterized by FTIR and XRD. The anti‐inflammatory and antioxidant activities were investigated in LPS‐stimulated RAW 264.7 macrophages using RT‐PCR and ELISA techniques for antioxidant‐ and inflammation‐related genes. The concentration of 39.6µg/ml of E.ang‐Fe0.25‐ZnO NPs demonstrated a significant anti‐inflammatory activity by suppressing the mRNA levels of TNF‐α and IL‐6 by 88.3%±1.9 and 93.6%±0.1, respectively, compared to LPS‐stimulated cells. This was confirmed by the significant reduction of TNF‐α and IL‐6 secretion levels from 95.2 and 495.6 pg/ml in LPS‐stimulated cells to 5.6 and 26.5 pg/ml in E.ang‐Fe0.25‐ZnO treated group. In addition, E.ang‐Fe0.25‐ZnO NPs nanoparticles treatment significantly enhanced the expression of antioxidant‐related genes, SOD and CAT. Together, our results proved that phyto‐mediated Fe0.25‐ZnO nanoparticles using Elaeagnus angustifolia have great potential in biomedical applications such as anti‐inflammatory and antioxidant.