干扰素刺激基因GPR135对水疱性口炎病毒复制的影响

Effects of interferon-stimulated gene GPR135 on the replication of vesicular stomatitis virus

目的探究G蛋白偶联受体(GPCR)135在病原体感染下的表达变化及其对水疱性口炎病毒(VSV)复制的影响.方法采用实时荧光定量聚合酶链式反应(Q-PCR)检测小鼠腹腔巨噬细胞(PEMs)中Gpr135在病原体感染和干扰素刺激下的表达变化.构建真核表达载体,在HEK-293T细胞中过表达GPR135.检测过表达GPR135对VSV复制的影响.结果在病原体感染PEM的情况下,Gpr135相对表达上调(P<0.05).直接用干扰素刺激PEM也会导致Gpr135相对表达上调(P<0.05).干扰Stat1表达后,VSV感染和干扰素刺激均不能上调Gpr135表达.在HEK-293T细胞中过表达GPR135后发现VSV的RNA相对复制量减少(P<0.05),并且VSV糖蛋白(VSV-G)的蛋白水平减少.结论在病原体感染的情况下,干扰素通过JAK-STAT信号通路上调Gpr135表达,可视为干扰素刺激基因.在HEK-293T细胞中过表达GPR135可以抑制VSV复制.

Objective To investigate the expression changes of G protein-coupled receptor(GPCR)135 in pathogen infection and its effects on the replication of vesicular stomatitis virus(VSV).Methods The expressions of GPR135 in mouse peritoneal macrophages(PEMs)in the cases of pathogen infection and interferon stimulation were detected with real-time fluorescent quantitative polymerase chain reaction(Q-PCR).The eukaryotic expression vector was constructed to overexpress GPR135 in HEK-293T cells.The effects of GPR135 overexpression on VSV replication were examined.Results In pathogen infection,GPR135 expression was increased(P<0.05).Interferon stimulation also led to an increase of GPR135 expression(P<0.05).After Stat1 interference,viral infection and interferon stimulation failed to upregulate Gpr135 expression.After GPR135 overexpression in HEK-293T cells,RNA replication of VSV was reduced(P<0.05),and expression of VSV glycoprotein(VSV-G)was significantly decreased.Conclusion In pathogen infection,interferon up-regulates GPR135 through the JAK-STAT signaling pathway and Gpr135 can be regarded as an interferon-stimulated gene.Overexpression of GPR135 in HEK-293T cells inhibits VSV replication.