调控Abi1基因的表达促进神经元轴突生长的实验研究  

Research on the correlation between the axon regeneration and Abi1 gene expression regulation

作　　者：黄忠会[1] 滕海军[2] 曹全斌[1] 

机构地区：[1]解放军第89医院急救中心,山东潍坊261200 [2]解放军第89医院脊柱外科,山东潍坊261200

出　　处：《中国急救医学》2008年第12期1095-1099,I0011,共6页Chinese Journal of Critical Care Medicine

摘　　要：目的探讨用RNA干扰技术抑制神经元细胞Abi1(非受体蛋白酪氨酸激酶Abl的结合蛋白)基因的表达对轴突生长的影响。方法设计四个针对Abi1的特异性小干扰RNA(siR-NA),应用体外转录合成siRNA。分别转染培养的神经元细胞,用半定量PCR及Western blot方法观察4段RNA对细胞内Abi1基因的表达影响,并选取具有最佳抑制效应的siRNA,转染神经元细胞,加入轴突生长抑制物,观测轴突生长情况。结果转染了4段siRNA的神经元细胞中Abi1mRNA和蛋白水平均明显降低,具有最佳抑制效应的片段是siRNA(325-345),转染最佳抑制效应片段的神经元细胞轴突能在轴突生长抑制物作用下继续生长。结论应用RNA干扰技术能高效抑制神经元内Abi1基因的表达,Abi1基因的表达下调促进了损伤神经元轴突的生长。Objective To invistigate the influence of RNA interference in cultured neurons ,and observe the effects of Abil gene silenceing on neurite regeneration. Methods Four Abil specific sirnas and a negative control siRNA were designed and then synthesized by in intro transcription, siRNA were transfected into cultured neurons respectively ,72 h later, the mRNA and protein expression of Abil was detected by semiquantitative RT - PCR and western blot analysis, after that, neurons transfected with the most efficient siRNA were cultured with myelin for 48 h, the neurite regeneration were invistigated. Results RNA interference can efficiently suppress Abil espression in cultured neurons, which to promote neurite regeneration. The mRNA and protein expression of Abil in neurons was significently down - regulated by 4 siRNA sequences. The most effective sequence was siRNA （ 325 - 345 ） , neurons transfected with it survived through myelin . Conclusions RNA interference can efficiently suppress Abil espression in cultured neurons,which to promote neurite regeneration.

关 键 词：调控 非受体蛋白酪氨酸激酶Ab1的结合蛋白(Abi1) 神经细胞 轴突 

分 类 号：R363[医药卫生—病理学]