IGFBP7在H1N1流感病毒引发的肺上皮细胞炎症和凋亡中的作用及机制

Role of IGFBP7 in H1N1 influenza virus-induced inflammation and apoptosis in lung epithelial cells and related mechanism

目的探究胰岛素样生长因子结合蛋白7(IGFBP7)在H1N1流感病毒引发的肺上皮细胞炎症和凋亡中的作用及机制。方法Western blot实验检测H1N1病毒对肺上皮A549细胞中IGFBP7表达的影响;MTT法检测A549细胞的活力;ELISA试剂盒检测炎症因子水平;Western blot实验检测凋亡相关蛋白cleaved-caspase-3的表达。结果H1N1病毒显著抑制A549细胞的活力且具有剂量、时间依赖性(P<0.05);H1N1病毒显著促进IGFBP7的表达且具有剂量、时间依赖性(P<0.05);干扰IGFBP7显著降低H1N1诱导的A549细胞的炎症因子水平(P<0.05);干扰IGFBP7显著抑制细胞外调节蛋白激酶(ERK)和p65磷酸化(P<0.05);过表达ERK或核因子κB(NF-κB)显著逆转IGFBP7对A549细胞炎症和凋亡的影响(P<0.05)。结论干扰IGFBP7的表达能够通过调控ERK/NF-κB信号通路减轻H1N1诱导的肺上皮A549细胞的炎症反应,并减少A549细胞的凋亡,为甲型流感病毒引发的肺损伤的治疗提供了新思路。

Objective To investigate the role of insulin-like growth factor-binding protein 7(IGFBP7)in H1N1 influenza virus-induced inflammation and apoptosis in lung epithelial cells and related mechanism.Methods The effect of H1N1 virus on IGFBP7 expression in A549 cells was detected by Western blot.The viability of lung epithelial A549 cells was assessed by MTT assay.The levels of inflammatory factors were examined by ELISA kits.The expression of apoptosis-associated protein cleaved-caspase-3 was measured by Western blot.Results H1N1 virus significantly inhibited the viability of A549 cells in a dose-time dependent manner(P<0.05).H1N1 virus markedly promoted IGFBP7 expression in a dose-time dependent manner(P<0.05).Interfering IGFBP7 dramatically reduced the levels of H1N1-induced inflammatory factors in A549 cells(P<0.05).Interfering IGFBP7 remarkably inhibited the phosphorylation of extracellular regulated protein kinase(ERK)and p65(P<0.05).Overexpression of ERK or nuclear factor κB(NF-κB)significantly reversed the effect of IGFBP7 on inflammation and apoptosis in A549 cells(P<0.05).Conclusions Interfering IGFBP7 attenuates H1N1-induced inflammatory response and apoptosis in A549 cells through regulating the ERK/NF-κB signaling pathway,which provides a new idea for the treatment of influenza A virus-induced lung injury.