AD中REST的空间分布对神经元细胞凋亡与自噬的影响

目的 探讨Alzheimer病中抑制性因素1沉默作用转录因子(repressor element-1 silencing transcription, REST)的空间分布对神经元的凋亡与自噬的影响。方法 C57小鼠侧脑室注射Aβ25-35建造AD模型,以无菌生理盐水为对照,用荧光共聚焦显微镜观测REST蛋白在造模前后细胞内的定位的变化,并采用Real-Time PCR和Western印迹法检测小鼠神经元中凋亡与自噬相关基因的表达。结果 荧光共聚焦显微镜观测发现AD状态下REST基因入核受阻,Real-Time PCR和Western印迹法显示促进凋亡的BAX、CASP9表达增强,而抑制凋亡的BCL2基因表达下降。自噬标志基因LC-3的表达水平也增强。结论 AD状态下REST基因入核受阻,并引起凋亡与自噬相关基因的表达。
Objective To investigate the effect of repressor element-1 silencing transcription(REST) spatial distribution on apoptosis and autophagy of neurons in mouse Alzheimer's disease(AD) model. Methods C57 mice were randomly divided into AD group and normal control group. Mouse AD model was established by injection of Aβ25-35 through lateral cerebral ventricle. The location of REST protein in cell was monitored by confocal microscope. The mRNA and protein expressions associated with neuronal apoptosis and autophagy were tested by Real-Time PCR and Western blotting, respectively. Results REST factor was blocked into nuclear in AD mice. Compared with normal group, the expression of pro-apoptotic genes BAX and CASP9 were increased significantly, while the expression of apoptosis-inhibiting gene BCL2 was decreased significantly. Meanwhile expression of autophagy gene LC-3 was enhanced. Conclusion REST protein is blocked for nuclear transfer in Aβ- induced mouse AD model, which may induce the changes of apoptosis and autophagy-related gene expression