To investigate the role of HIF-1α in inflammation mediated by macrophage infect with H1N1(PR8). Mouse macrophages(RAW264.7) were infected with PR8 and sacrificed at 0 h, 6 h, 12 h, 24 h, 48 h post infection(HPI), the mRNA expression levels of HIF-1α， IL-6, TNF-α， M protein(MBP) were determined by real-time PCR. HIF-1α and NF-κB, MAPK, Akt, MBP protein expression levels were detected by Western Blot. In inhibition test, HIF-1α was inhibited by 2-MeOE-2(10 nmol/L), after 24 h, IL-6, TNF-α， MBP mRNA levels and NF-κB, MAPK, Akt, MBP NF-κB, MAPK, Akt protein levels were detected. The results demonstrated that the viral load in infected cells peaked on 24 HPI, HIF-1α mRNA level increased transiently 6 HPI, and peaked on 24 DPI，so it is the same with IFN-γ、IL-6、TNF-α mRNA. After inhibiting HIF-1α in PR8-infected RAW264.7 cells with 2-MeOE-2(10 nmol/L), IL-6、TNF-α mRNA level were decreased when compared with control group (P<0.05). NF-κB protein expression was also decreased when compared with control group. It was concluded that HIF-1α may induce inflammation in H1N1 infection through up-regulating NF-κB pathway of macrophage.