H9N2亚型流感病毒感染过程中神经介素B及受体对NF-κB信号通路泛素酶的调控

Ubiquitinase of NF-κB Signal Pathway Regulated by Neuromedin B and Its Receptor NMBR during Influenza A Virus H9N2 Subtype Infection

神经介素B(neuromedin B,NMB)及受体(NMB receptor,NMBR)通过NF-κB信号通路参与抑制甲型流感病毒(influenza A virus,IAV)H1N1亚型(IAV/H1N1)的感染。但关于NMB和NMBR对NF-κB信号通路相关泛素蛋白酶的调控如何,尚未见报道。为探究NMB和NMBR调控IAV诱导的NF-κB信号通路相关的泛素蛋白酶表达的影响,本研究基于IAV/H9N2感染sh-NMBR细胞与NMB刺激的A549细胞,采用RT-PCR、qRT-PCR及Western blot(WB)分析NMB和NMBR对H9N2感染引起的NF-κB信号通路相关的E3泛素连接酶Mind bomb-2(MIB2)和Ring Finger Protein 8(RNF8)及去泛素蛋白酶Cylindromatosis(CYLD)的表达变化。结果显示:H9N2感染sh-NMBR细胞中,RNF8和CYLD表达增加,MIB2表达和P65磷酸化水平降低;NMB激活A549细胞中NMBR的表达后,诱导细胞中RNF8和CYLD的表达水平下降,MIB2表达和P65磷酸化水平增加。结果表明:NMB和NMBR通过调控IAV/H9N2感染诱导的泛素蛋白酶的表达和P65活性,从而影响IAV/H9N2感染激活的NF-κB信号通路的功能,该结果为深入研究NMB和NMBR抑制甲型流感病毒感染的作用机制提供了理论基础。

Neuromedin B(NMB)and its receptor(NMBR)could inhibit the infection of influenza A virus(IAV)H1N1 subtype through activating the pathway of NF-κB signaling.However,the regulation of NMB and NMBR on the expression of ubiquitination ligases related with NF-κB signaling pathway remains unclear.To explore the effects of NMB and NMBR on regulating the ubiquitination ligases involved in NF-κB signaling pathway during IAV/H9N2 infection,the expression levels of E3 ubiquitin ligase mind bomb-2(MIB 2)and ring finger protein 8(RNF 8),deubiquitin enzyme cylindromatosis(CYLD)involved in NF-κB signaling pathway were analyzed using the methods of RT-PCR,qRT-PCR,and Western blot(WB),based on the sh-NMBR cells and NMB stimulating A549 cells during IAV/H9N2 infection.The results showed that the increased expression levels of RNF8 and CYLD,and the decreased levels of MIB 2 and P65 phosphorylation were observed in sh-NMBR cells during H9N2 infection,while the decreased levels of RNF8 and CYLD,and the increased levels of MIB 2 and P65 phosphorylation were confirmed in NMB stimulating A549 cells.These results indicated that NMB and NMBR could affect the function of NF-κB signaling pathway by regulating the expression levels of ubiquitination ligases and P65 phosphorylation during IAV/H9N2 infection,which could provide a theoretical basis for further studying the mechanism of NMB and NMBR on inhibiting influenza A virus.