Because it is highly contagious, the influenza A virus (IAV) has the potential to cause pandemics in humans. The emergence of drug-resistant strains requires the development of new chemical therapeutics. Oroxylin A (OA) is a flavonoid which has been shown to have antioxidant and antitumor effects. However, intensive studies in which OA fights against different influenza virus strains and the underlying antiviral mechanisms have not been reported. In our study, the antiviral activities in cells and in mice, the preliminary mechanisms of OA were investigated. Our data show that it can inhibit A/FM/1/47 (H1N1), A/Beijing/32/92 (H3N2) and oseltamivir-resistant A/FM/1/47-H275Y (H1N1-H275Y) viruses in MDCK cells in a dose-dependent manner with inhibitory rates of 70.9%, 59.5% and 23.2%, respectively, at 50μM doses. Orally administered OA effectively protected mice from H1N1 virus-induced death, body weight loss and lung injury, with a survival rate of 60.0% at 100mg/kg/d dose. In addition, the H1N1 M1 gene transcription and protein synthesis were suppressed by 43.7% and 33.2%, respectively, in the late biosynthesis stage. This resulted in inhibition of viral replication. Furthermore, we found that OA has a neuraminidase (NA) inhibitory effect with IC₅₀ values for H1N1-H275Y and A/Anhui/1/2013-R294K (H7N9-R294K) of 241.4μM and 203.6μM, respectively. Interferons (IFNs) produced by the virally infected cells play important roles in antiviral defense, therefore, IFN levels in the blood were also tested in mice. We found that IFN-β and IFN-γ in the OA 100mg/kg/d group were markedly increased by 24.5pg/mL and 859.9pg/mL, respectively, compared with those in the model group. This indicated that OA could induce the secretion of IFNs. The potent inhibition of virus replication and NA inhibitory activity, as well as the promotion of IFN production suggest that OA could be a drug candidate to fight against IAVs including oseltamivir-resistant strains.