冷诱导RNA结合蛋白通过激活NF-κB信号通路影响H1N1甲型流感病毒的复制

doi:10.11843/j.issn.0366-6964.2016.10.020

摘要/Abstract

摘要：

冷诱导RNA结合蛋白（CIRP）是NF-κB和ERK信号通路的上游调控因子，而这两条信号通路是甲型流感病毒复制和机体起始免疫所必需的，为探讨CIRP对H1N1甲型流感病毒复制的影响及可能的分子机制，构建了CIRP过表达BHK-21细胞系（Cirp+BHK-21），用Western blot检测NF-кB和ERK1/2的磷酸化水平，研究CIRP对NF-кB和ERK1/2的调节作用；用Real-time RT-PCR检测H1N1甲型流感病毒感染后Cirp+BHK-21和对照细胞中病毒拷贝数的动态变化，以及在特异性阻断剂PDTC阻断NF-кB通路的Cirp+BHK-21细胞中病毒拷贝数的动态变化。Western blot检测结果显示：过表达CIRP显著促进了BHK-21细胞中NF-κB的磷酸化水平（P<0.05），而对ERK1/2的磷酸化水平无显著影响；病毒定量检测结果显示：过表达CIRP能显著促进H1N1甲型流感病毒的增殖，感染后3、9、15、21 h 病毒在Cirp+BHK-21细胞中的拷贝数分别为对照组的111%、103%、167%和235% (P<0.05）；阻断NF-κB信号通路后病毒的拷贝数显著下降，在感染后3、9、15、21 h分别为未阻断组的98%、42%、19%（P<0.05）和7%（P<0.05）。从本研究结果可见，CIRP可通过活化NF-κB信号通路促进H1N1甲型流感病毒的复制。

Abstract:

The cold-inducible RNA-binding protein (CIRP) is an upstream regulator of the NF-κB and ERK pathways，which are essential for the replication of the influenza virus and the initial immune response.In order to investigate the effect of CIRP on the replication of H1N1 influenza A virus and its possible molecular mechanism，in this study，CIRP overexpression BHK-21 (Cirp+BHK-21) cells were constructed，and the phosphorylation levels of NF-κB and ERK in Cirp+BHK-21 cells were detected by Western blot to confirm the effect of CIRP on regulation of NF-κB and ERK1/2；Real-time RT-PCR was used to detect the dynamic changes of virus load in Cirp+BHK-21 and control cells after infected with influenza A virus，the method was also used to detect the dynamic changes of virus load in Cirp+BHK-21 cells which were blocked by the NF-κB inhibitor PDTC.The results of Western blot exhibited that overexpressed CIRP could significantly increase the expression of phosphorylation level of NF-κB (P<0.05)，but had no significant effect on the phosphorylation level of ERK.The results of quantitative detection of virus showed that overexpressed CIRP could significantly enhance the proliferation of influenza A virus，the virus load in the Cirp+BHK-21 cells were 111%，103%，167% and 235% (P<0.05) at 3，9，15 and 21 h PI，respectively，compared to the control group；Blocking the NF-κB was significantly decreased the virus load in the Cirp+BHK-21，and the virus load in treatment group were 98%，42%，19% (P<0.05)，7% (P<0.05) at 3，9，15 and 21 h PI，respectively，compared to the unblock group.Therefore，this study confirmed that overexpressed CIRP could enhance the proliferation of influenza A virus via activation of NF-κB pathway.

中图分类号:

S852.23

聂培婷，汤承，岳华. 冷诱导RNA结合蛋白通过激活NF-κB信号通路影响H1N1甲型流感病毒的复制[J]. 畜牧兽医学报, 2016, 47(10): 2108-2114.

NIE Pei-ting，TANG Cheng，YUE Hua. Activation of Cold-inducible RNA-binding Protein by H1N1 Influenza Virus Contributes to Viral Replication Via Activating NF-κB Pathway[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(10): 2108-2114.

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