Update publication information. Highly pathogenic avian influenza virus (HPAIV) is a high morbidity-and mortality-associated disease that causes rapid systemic illness and death in susceptible animals. Lycorine, as the major component of Amaryllidaceae alkaloids, exhibits better protective effects against A/Chicken/Guangdong/178/2004 (GD178) than the commercial neuraminidase (NA) inhibitor oseltamivir in our prior study. Although the change of proteins of the whole cell in response to HPAIV infection have been revealed by proteomics, the mechanisms resulted in lycorine-related symptoms and the crucial factors remain to be elucidated. In this study, we carried out Multiplex tandem mass tag (TMTs) approach to analyze protein level variation of virus-infected host cells treated by lycorine using. Three groups were administrated: mock infection group (L), virus infection group (M) and virus infection and lycorine-treated after virus infection group (H). 5786 proteins were identified, among which from these three groups using TMT proteomics analysis. In M/L group, 1101 proteins were identified, of which 340 proteins were determined differentially expressed proteins during HPAIV; meanwhile, 1059 proteins were identified from lycorine-treated group, among which 259 proteins presented significant change. Here, proteins in M/L and M/H were acquired simultaneously trend, and the proteins in each fraction were analyzed. Through the enrichment analysis of Gene Ontology (GO)and KEGG pathways, we found that the up-regulated proteins in the mitogen-activated protein kinase signaling pathway and RNA transport were most enriched after HPAIV infected. There were 70 differentially abundant proteins with co-changed proteins in M/L and M/H groups were selected as the candidate proteins. Moreover, we noticed that the nuclear pore complex protein (Nup) 93 was associated with both mitogen-activated protein kinase signaling pathway and RNA transport. In addition, the Western blot experiments confirmed that the expression of Nup93 was significantly down-regulated in lycorine treatment but induced after viral infection. Conclusion: Thus, the research may provide new insight into lycorine’s underlying anti-influenza mechanisms and potential therapeutic targets for influenza virus