Ephedrannin B (EPB) has been shown to exert anti-inflammatory effects. However, the effect of EPB onrespiratory syncytial virus infection (RSV) is not known. In this study, the cytotoxic effect of EPB wasevaluated in BEAS-2B cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Reversetranscription quantitative polymerase chain reaction and Western blot assays were performed to determinethe expression of target genes. The anti-viral effect of EPB was assessed by determining viral titers usingplaque assay. We found that RSV infection caused a marked increase in interleukin (IL)-6, IL-8, IL-1beta andtumor necrosis factor (TNF)-alpha production and release, which was concentration-dependently attenuated byEPB treatment. Furthermore, EPB decreased the expression of RSV fusion gene in RSV-infected BEAS-2Bcells. Concomitantly, EPB treatment led to an obvious inhibition of viral replication in BEAS-2B cells.Besides, EPB suppressed RSV-induced mitogen-activated protein kinase/nuclear factor kappa-light-chainenhancerof activated B cells signaling. In conclusion, EPB exerts anti-viral and anti-inflammatory propertiesin BEAS-2B cells infected with RSV.