【目的】 探讨安石榴苷抗甲型 H1N1流感病毒感染的作用及机制。【方法】 将 60只 BALB/c雄性小鼠分为正常组，模型 组，达菲组及安石榴苷低、中、高剂量组，每组20只。除正常组，其他各组小鼠建立甲型H1N1流感病毒感染模型。连续给 予相应药物4 d。检测小鼠体质量，器官（脾脏、肺脏）指数及其抑制率，肺脏组织流感病毒载量，14 d内存活率。苏木精-伊 红（HE）染色法观察小鼠肺脏组织病理变化。酶联免疫吸附分析（ELISA）检测小鼠脾脏组织中细胞因子肿瘤坏死因子α（TNF-α）、 白细胞介素（IL）-6、IL-17、干扰素γ（IFN-γ）水平；蛋白免疫印迹（Western Blot）法检测小鼠脾脏组织中磷酸化p38丝裂原活 化蛋白激酶（p-p38MAPK）、磷酸化细胞外信号调节激酶（p-ERK）1、p-ERK2蛋白表达量。【结果】 模型组、达菲组和安石榴 苷低、中、高剂量组的 14 d内存活率分别为 10%、80%、50%、70%、80%。与正常组比较，模型组小鼠体质量、14 d内存 活率显著降低（P＜0.05），肺脏和脾脏指数，肺组织流感病毒载量，TNF-α、IL-6水平及p-p38MAPK、p-ERK1、p-ERK2蛋 白表达量显著升高（P＜0.05），HE染色显示肺组织间质有明显的炎性细胞浸润，肺泡间隔增厚。与模型组比较，达菲组和安 石榴苷中、高剂量组小鼠体质量，14 d内存活率，脾脏组织IL-17、IFN-γ水平显著升高（P＜0.05），肺脏和脾脏指数，肺组 织流感病毒载量，TNF-α、IL-6水平及p-p38MAPK、p-ERK1、p-ERK 2蛋白表达量显著降低（P＜0.05），HE染色可见肺组 织病理变化有所改善。达菲组和安石榴苷低、中、高剂量组的肺脏指数抑制率分别为 42.75%、20.19%、36.74%、42.29%， 脾脏指数抑制率分别为 43.97%、29.66%、47.80%、53.68%。【结论】 安石榴苷可通过抑制 MAPK/ERK 信号通路减轻甲型 H1N1流感病毒感染小鼠肺部损伤，降低病毒载量，调节关键细胞因子，抑制炎症反应，从而发挥免疫调节作用，抵抗甲型 H1N1流感病毒，提高存活率。

Objective To elucidate the impact of punicalagin on a mouse model infected with influenza A H1N1 virus and to investigate its immunomodulatory mechanism based on the mitogen-activated protein kinase （MAPK）/ extracellular signal-regulated kinase （ERK） signaling pathway. Methods Sixty BALB/c male mice were divided into a control group， a model group， an oseltamivir group， and low-dose punicalagin group， medium-dose punicalagin group，and high-dose punicalagin group，with 20 mice in each group. The corresponding drugs were administered continuously for 4 days. The body mass，organ （spleen and lung） indices and inhibition rates，viral load in lung tissue，and survival rate within 14 days were measured. Histopathological changes in lung tissue were observed using hematoxylin and eosin （HE） staining. The levels of cytokines tumour necrosis factor-α（TNF-α）， interleukin （IL）-6， IL-17， and interferon- γ（IFN- γ） in spleen tissue were detected using enzyme-linked immunosorbent assay （ELISA） kits. Protein levels of phospho （p）-p38MAPK，p-ERK1，and p-ERK2 in spleen tissue were assessed using Western Blot. Results The 14-day survival rates of the model group，oseltamivir group， and low- ， medium- ， and high-dose punicalagin groups were 10%， 80%， 50%， 70%， and 80%， respectively. Compared with the normal group，the model group showed significantly decreased body mass and 14- day survival rate （P＜0.05），while lung and spleen indices，influenza viral load in lung tissue，levels of TNF-α and IL-6，and protein expression levels of p-p38MAPK，p-ERK1，and p-ERK2 were significantly increased （P＜0.05）. HE staining revealed significant inflammatory cell infiltration in lung interstitium and thickened alveolar septa. Compared with the model group，the oseltamivir group and medium- and high-dose punicalagin groups showed significantly increased body mass，14-day survival rate，and levels of IL-17 and IFN-γ in spleen tissue （P＜0.05），while lung and spleen indices，influenza viral load in lung tissue，levels of TNF-α and IL-6， and protein expression levels of p-p38MAPK，p-ERK1，and p-ERK2 were significantly decreased （P＜0.05）. HE staining showed improved pathological changes in lung tissue. The lung index inhibition rates of the oseltamivir group and low-，medium-，and high-dose punicalagin groups were 42.75%，20.19%，36.74%，and 42.29%， respectively， while the spleen index inhibition rates were 43.97%， 29.66%， 47.80%， and 53.68%， respectively. Conclusion Punicalagin can alleviate lung injury in mice infected with influenza A H1N1 virus， reduce viral load， regulate key cytokines， and suppress inflammatory responses by inhibiting the MAPK/ERK signaling pathway， thereby exerting immunomodulatory effects against influenza A H1N1 virus and improving survival rate.

R285.5

山西省应用基础研究计划项目（编号：201901D111393）