摘要
目的 mir-153可负调控阿尔茨海默病(Alzheimer’s disease,AD)主要致病基因APP及APLP2的蛋白表达,降低其胞内降解片段(intracellular domains,ICDs)的生成。因ICDs具有转录活化及促凋亡活性,本研究旨在探讨mir-153对这两个靶基因下游信号分子GSK-3β表达水平及细胞抗损伤能力的影响,以期进一步阐明mir-153在阿尔茨海默病发病机制中的作用。方法构建mir-153稳转细胞系及mir-153转基因小鼠,Western blot检测该细胞系及小鼠脑内磷酸化GSK-3β、Tau及其总蛋白的表达;Aβ42肽和H2O2分别处理mir-153稳转细胞系,MTS法检测细胞增殖活性的改变,流式细胞术检测细胞凋亡水平的改变。结果 mir-153稳转细胞系中磷酸化GSK-3β及其总蛋白的表达下调,Tau磷酸化水平降低。mir-153转基因小鼠脑内,磷酸化GSK-3β及其总蛋白的表达降低,磷酸化Tau及其总蛋白水平均无明显变化。Aβ42肽和H2O2损伤作用下,mir-153稳转细胞系的增殖活性显著降低,凋亡水平增加。结论 mir-153可负调控靶基因下游信号分子GSK-3β的表达;高表达mir-153可降低细胞抗损伤的能力。
Objective Mir-153 can negatively regulate the expression of APP and APLP2 protein,the crucial Alzheimer＇s disease related genes,and consequently lower the level of their intracellular degradation fragment（intracellular domains,ICDs）.Considering the transcriptional activity and pro-apoptotic role of ICDs,the aim of this study was to explore the effect of mir-153 on the expression of GSK-3β,the downstream signaling molecule of the two target genes,and on the ability of cells against damage stress to further identify the role of mir-153 in Alzheimer＇s disease.Methods A stably transfected cell line over-expressing mir-153 was developed and mir-153 transgenic mice were generated.Western blot was used to detect the expression of phosphorylated GSK-3β,Tau and their total protein in the cells and mice.The mir-153 stably transfected cells were treated with Aβ42 peptide and H2O2,respectively,to determine the changes of cell viability by MTS and analyze the cell apoptosis by flow cytometry.Results The expression of phosphorylated GSK-3β and it＇s total protein were decreased and the phosphorylation of Tau was reduced in the mir-153 stably transfected cells.The expression of phosphorylated GSK-3β and it＇s total protein were down-regulated and the level of phosphorylated Tau and its total protein were not significantly changed in the brain of mir-153 transgenic mice.Under the treatment of Aβ42 peptide and H2O2,the viability of mir-153 stably transfected cells were clearly decreased and the apoptosis level of the cells was increased.Conclusion Mir-153 can negatively regulate the expression of GSK-3β,the downstream signaling molecule of its target genes.Over-expressed mir-153 lowers the cellular anti-injury ability.
出处
《中国比较医学杂志》
CAS
2011年第8期15-19,共5页
Chinese Journal of Comparative Medicine
基金
卫生部科研专项项目(200802036)
