狂犬病病毒(RABV)感染小鼠原代神经元细胞对Bif-1蛋白表达的影响

Effect of rabies virus infection on expression of Bif-1protein in primary neurons of mice

建立狂犬病病毒(rabies virus,RABV)CVS-11株感染小鼠原代神经元细胞模型,探究RABV感染小鼠原代神经元细胞对Bif-1(Bax-interacting factor-1)蛋白表达的影响。分离并培养小鼠大脑皮质原代神经元细胞,通过间接免疫荧光鉴定RABV感染效率。将RABV以MOI=0.1剂量接种原代神经元细胞,使用直接免疫荧光、Western blot、RT-PCR等方法检测RABV在原代神经元细胞中的复制情况。利用Western blot检测RABV感染原代神经元细胞后Bif-1蛋白的表达情况。结果成功分离了小鼠大脑皮质原代神经元细胞;直接免疫荧光、Western blot、RT-PCR结果显示,RABV CVS-11株可感染原代神经元细胞;Western blot结果表明RABV感染后,原代神经元细胞中Bif-1蛋白水平早期出现短期上升后降低,且整体低于正常水平。RABV CVS-11感染原代神经元细胞后,Bif-1蛋白水平发生改变,为进一步探索Bif-1蛋白在RABV致病中的作用及其分子机制奠定基础。

To establish a model of rabies virus CVS-11infection in mouse primary neuronal cells and to explore the effect of rabies virus infection on the expression of Bif-1(Bax-interacting factor-1)protein in mouse primary neuronal cells,mouse cerebral cortex primary neuronal cells were isolated,cultured and infected with rabies virus at a dose of MOI=0.1,and direct immunofluorescence,Western blot and RT-PCR were used to detect the replication of rabies virus in primary neuronal cells.The expression of Bif-1protein in primary neuronal cells after rabies virus infection was detected by Western blot.The primary neuronal cells of the mouse cerebral cortex were successfully prepared.The results of direct immunofluorescence,Western blot,and RT-PCR showed that the rabies virus CVS11strain could infect and replicate in primary neuronal cells.The Western blot results showed that after rabies virus infection,the expression level of Bif-1protein in primary neuron cells increased for a short period of time in the early stage and then gradually decreased,and overall level was lower than normal.Our study lay a foundation for further study on the relationship between Bif-1protein and the pathogenicity of rabies virus.