慢病毒介导的CBS基因过表达对原代皮质神经元β淀粉样蛋白分泌的影响

Effect of Overexpression of CBS Mediated by Lentivirus on Aβ in Cortical Neurons

目的通过过表达手段上调胱硫醚β-合成酶(Cystathionine-β-synthase,CBS)在原代神经元细胞中的表达,观察其对原代神经元β淀粉样蛋白(Aβ)分泌的影响。方法使用过表达CBS慢病毒载体感染体外培养的原代皮质神经元,设定为慢病毒介导阳性(CBS)组、另设阴性对照慢病毒感染(NC)组、未经慢病毒感染(CON)组。采用实时荧光定量PCR检测CBS基因mRNA的表达,Western blot检测CBS蛋白的表达,采用Elisa检测Aβ40的表达水平。结果使用过表达CBS的慢病毒载体感染体外培养的神经元,实时荧光定量PCR检测其mRNA的表达上调明显升高,与NC组和CON组相比差异有统计学意义(P<0.01),Western blot结果显示蛋白表达升高与定量PCR一致,Elisa检测感染72小时后CBS组、NC组、CON组Aβ40的分泌分别为(182.19±24.52)ng/L、(411.90±12.43)ng/L、(390.41±54.53)ng/L,CBS组与NC组和CON组相比差异有统计学意义(P<0.05)。结论慢病毒介导的CBS基因过表达可降低原代皮质神经元Aβ40的分泌。

Objective To over-express Cystathionine-β-Synthase（CBS）gene,and investigate its effect on Aβin cortical neu-rons.Methods The cortical neurons were infected with the packaged lentivirus,and it was designated as CBS group.Cells infected with CBS-NC were designated as NC group and not-infected cell was CON group.The levels of CBS mRNA were de-tected by real-time quantitative PCR.The CBS protein was detected by western blot and the levels of Aβ40 were measured by ELISA（enzyme-linked immunosorbent assay）.Results After the infection of CBS mRNA recombinant lentivirus .the ex-pression level of CBS mRNA in the cortical neurons increased significantly versus NC and CON groups（P〈0.01）.The up-dated expression of CBS proteins was consistent with CBS mRNA .The levels of Aβ40 were （182.19 ±24.52 ）ng/L、（411.90 ±12.43）ng/L and（390.41 ±54.53）ng/L in CBS,NC and CON groups respectively.As compared with NC and CON groups.Conclusion The overexpression of CBS Mediated by Lentivirus mRNA effectively increases the levels of Aβ40 in the cortical neurons.