Objective:To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo.Methods:In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase assays in Madin Darby canine kidney cells.In vivo anti-influenza virus assays were performed on the viral pneumonia model of mice.The numbers of mice that died within day 2 to day 14 postinfection were recorded to calculate the mortality.On days 2,4,and 6,the viral titers in the lungs were determined by hemagglutination assay;hematoxylin/eosin staining was used to assess the pathogenic changes of lung tissues; the concentrations of tumor necrosis factor-alpha(TNF-α) and monocyte specific chemoattractant molecule (MCP-1) were measured by radio immunoassay or enzyme-linked immunosorbent assay;the concentrations of nitric oxide(NO) and inducible nitric oxide synthetase(iNOS) were detected by colorimetric method;reverse transcription polymerase chain reaction was used to detect the mRNA level of TNF-αand MCP-1.Results: Berberine showed inhibitory effects on cytopathogenic effects and neuraminidase activity of virus,with the therapeutic index 9.69.In vivo,berberine decreased mice mortality from 90%to 55%,reduced virus titers in the lungs on day 2 postinfection(P0.05).The lung histology scores were 1.50±0.67,4.50±1.00,and 5.50±1.00 in the berberine group on days 2,4,and 6,respectively,which were significantly reduced compared to 2.17±0.22,6.83±0.44,and 8.50±0.33 in the infected group(P0.05).The productions of NO and iNOS were repressed by berberine compared with those in the infected group(P0.01).The transcription and expression of TNF-αwere inhibited by berberine on day 4(P0.01) and day 6(P0.05),and those of MCP-1 were inhibited on day 6(P0.01) compared with the infected group.Conclusions:Berberine exhibited antiviral effects on the influenza virus both in vitro and in vivo.The possible therapeutic mechanism of berberine on influenza-induced viral pneu