过表达miR-132改善阿尔茨海默病样学习记忆障碍  

Overexpression of miR-132 attenuates Alzheimer disease-like learning and memory impairment

作　　者：王雪银 李宜培 程相树[2] 赵昆朋[2] 

机构地区：[1]河南医学高等专科学校,河南郑州451191 [2]河南大学医学院,河南开封475004

出　　处：《中国病理生理杂志》2016年第1期41-45,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81401062;No.U1504809)

摘　　要：目的:获得过表达微小RNA-132(microRNA-132,miR-132)的慢病毒,并检测对Tg2576小鼠学习记忆损伤的改善。方法:从小鼠脑组织中克隆miR-132成熟序列上下游各100 bp的基因,将基因克隆至慢病毒表达载体p Lenti7.3,酶切鉴定并且DNA序列测定正确。p Lenti7.3-miR-132和Vira Power Packaging Mix共转染293FT细胞。72 h后收获上请,采用超速离心和聚乙二醇法纯化病毒颗粒并分装保存。分别以病毒颗粒感染细胞系和小鼠海马组织验证病毒的感染能力。Tg2576小鼠海马区注射病毒,Western blot和Morris水迷宫分别检测记忆相关蛋白水平和空间学习记忆能力。结果:所包装的miR-132病毒颗粒能够高效地感染细胞系,并且在体内也有理想的感染效率。和对照组相比,过表达miR-132的Tg2576小鼠PSD95和GluR1明显升高,并且空间依赖的学习记忆能力明显提高。结论:过表达miR-132通过影响突触相关蛋白PSD95和GluR1的水平改善Tg2576小鼠的空间学习记忆能力障碍。AIM： To obtain miR-132 lentivirus and to detect the effect of miR-132 overexpression on the improvement of learning and memory impairment in Tg2576 mice. METHODS： The pri-miR-132 gene was purified from mouse brain and cloned into lentivirus vector p Lenti7. 3-miR-132. The miR-132 lentivirus was packaged by co-transfected p Lenti7. 3-miR-132 with Vira Power Packaging Mix into 293 FT cells. The miR-132 lentivirus was purified 72 h later from the supernatant by ultracentrifugation and poly（ ethylene glycol） method. The efficiency of miR-132 lentivirus was identified by transfection into N2 A cells and mouse hippocampus region. Tg2576 mice were injected with miR-132 virus in the hippocampus region and the expression of learning- and memory-related proteins and functions were determined. RESULTS：miR-132 lentivirus expression vector was constructed successfully. The purified virus was obtained with transfection activity. Tg2576 mice with overexpression of miR-132 had a significant improvement in learning and memory abilities. CONCLUSION： miR-132 lentivirus with biological activity was obtained. Overexpression of miR-132 in the hippocampus region of Tg2576 mice attenuates the learning and memory impairment.

关 键 词：微小RNA-132 病毒包装 Tg2576小鼠 学习 记忆 

分 类 号：R363.14[医药卫生—病理学]