为探索缺氧诱导因子(hypoxia inducible factor，HIF)-1α诱导甲型流感病毒毒株感染小鼠巨噬细胞引起炎症反应的具体机制，本研究以甲型H1N1流感病毒(简称H1N1)株A/PR/8感染小鼠巨噬细胞RAW264.7后，在显微镜下观察其在感染后的表型变化，分别在不同时间段收集样本，通过聚合酶链反应(polymerase chain reaction，PCR)检测HIF-1α、干扰素(interferon，IFN)-γ、白细胞介素(interleukin，IL)-6、肿瘤坏死因子(tumor necrosis factor，TNF)-α和M蛋白(M protein，MBP)mRNA的变化，通过蛋白质印迹法(Western blot, WB)检测HIF-1α、核转录因子-κB(nuclear factor-κB，NF-κB)、促分裂原活化的蛋白激酶(mitogenactivated protein kinase，MAPK)、蛋白激酶B(protein kinase B，Akt)以及M蛋白的变化。随后，加入抑制剂2-MeOE-2(10 nmol/L)进行抑制试验，采用PCR和WB检测HIF-1α表达被抑制后上述炎症因子mRNA表达水平及炎症蛋白通路的变化。结果显示，H1N1PR8感染小鼠巨噬细胞RAW264.7后，H1N1PR8复制率在24 h达到峰值，HIF-1α mRNA在感染6 h后开始升高，12 h迅速上升，24 h达到峰值。IFN-γ、IL-6、TNF-α mRNA变化趋势基本与HIF-1α一致，但在感染12 h并未进入快速上升期。HIF-1α蛋白在感染后6 h表达明显增多，24 h达到峰值，与mRNA变化水平基本一致。NF-κB通路蛋白在感染12 h后明显增多，48 h开始减少。加入抑制剂2-MeOE-2后，培养感染细胞24 h，抑制剂组IL-6、TNF-α mRNA水平较对照组显著下降(P<0.05)，抑制剂组NF-κB通路蛋白较对照组表达下降。本研究结果表明，小鼠巨噬细胞被H1N1感染后，HIF-1α可能通过激活NF-κB通路促进IL-6、TNF-α等炎症因子的分泌参与炎症反应。

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.