The study aims to clarify the therapeutic effect and underlying mechanisms of wogonin (WOG) against acute lung injury (ALI). Lipopolysaccharide-induced in vivo ALI mice model and in vitro RAW264.7 cell model were established. Lung histopathologic changes were assessed by H&E staining. Inflammatory cell infiltration in lung tissues and bronchoalveolar lavage fluid (BALF) was determined using H&E staining and flow cytometry. Inflammatory cytokines, reactive oxygen species (ROS), and antioxidant mediators were quantified. Potential targets of WOG were identified by bioinformatics and network pharmacology and further verified by Western blotting. WOG treatment (10 mg/kg, 20 mg/kg, 40 mg/kg) alleviated lung injury of ALI mice by ameliorating infiltration of inflammatory cells in lung tissues and BALF, inhibiting inflammatory cytokines, elevating antioxidant mediator levels, and attenuating lung edema. In RAW264.7 cells, WOG treatment (10 μM, 20 μM, 40 μM) also suppressed inflammatory response and oxidative stress. Peroxisome proliferator-activated receptor α (PPARα), protein kinase B (AKT), and nuclear factor erythroid 2-related factor 2 (NRF2) were predicted to be the targets of WOG against ALI by integrating the network pharmacology database and the GSE1871 dataset. Interestingly, WOG markedly promoted the expression and activation of PPARα and NRF2, while it inhibited the expression and phosphorylation of AKT both in vivo and in vitro. We provided a comprehensive insight into investigating the therapeutic effect of WOG against ALI via ameliorating inflammation and oxidative stress through regulating PPARα, AKT, and NRF2.