Better effective therapy for highly pathogenic influenza virus infection should consist of combinations of an effective antiviral agent and immunomodulatory agents to control viral replication and pulmonary injury, respectively. Here we evaluated the virus titres in different cells (A549 cells and MDCK cells) infected with influenza virus H5N1 by plaque assay with the various concentrations of Geldanamycin at different time points. The levels of proinflammatory cytokines from A549 cell infected with influenza virus H5N1were detected by ELISA in the absence or presence of Geldanamycin. Our findings showed that Geldanamycin significantly inhibited influenza virus growth in MDCK cells and A549 cells(P < 0.01). However, influenza virus growth were not inhibited with Geldanamycin on 36 h and 48 h(P > 0.05)after infection with influenza virus H5N1. Analysis of proinflammatory cytokines in Geldanamycin-treated A549 cells revealed significant reductions in IFN-α，TNF-α or IL-6 on 12 h and 24 h after infection with influenza virus H5N1(P < 0.05). All the results indicated that Geldanamycin have the dual effect of combine antivirus with anti-inflammation, which provided the science data for the potential use of Geldanamycin as a novel, highly effective anti-influenza virus drug.