摘要
目的研究β-淀粉样蛋白25-35(Aβ25-35)对大鼠骨髓间充质干细胞(BMSCs)分化的神经细胞Tau蛋白磷酸化的影响。方法体外分离纯化大鼠骨髓MSCs,将P4 MSCs分为两组:(1)Aβ25-35实验组中加入预诱导培养基(含10μg/L bFGF,10%FBS的DMEM)和20μmol/L Aβ25-35,24h后用含2%DMSO和200μmol/L BHA的DMEM诱导其向神经细胞分化,5h后收取细胞;(2)对照组诱导方法同Aβ实验组,但在诱导过程中未加入Aβ25-35。光镜下观察细胞形态变化,免疫细胞化学法检测神经神经元特异性烯醇化酶(NSE),western blotting检测GSK-3β、Tau[pSer262]和Tau[pSer396]。结果光镜下可见纺锤状的MSCs诱导后出现长突起,呈现神经细胞样形态,但Aβ25-35实验组突起数量和长度均少于对照组;免疫细胞化学法检测两组诱导后的神经元样细胞为NSE阳性;免疫蛋白印记法证明Aβ25-35实验组的GSK-3β、Tau[pSer262]和Tau[pSer396]表达均显著高于对照组。结论 Aβ25-35能够通过GSK-3β途径诱导骨髓间充质干细胞分化的神经样细胞Tau蛋白过度磷酸化。
Objective To study the effect of β-amyloid 25-35（Aβ25-35）on Tau protein phosphorylation of rat bone marrow mesenchymal stem cells（MSCs）differentiated neuron-like cells.Methods BMSCs were isolated and purified from SD rats,and the P4 BMSCs were divided into two groups：（1）Aβ25-35 experiment group：added with pre-induction medium（containing 10 μg/L bFGF,10%FBS in DMEM）and 20μmol/L Aβ25-35,24 hours later,the cells were induced into neurons-like cells by DMEM with 2%DMSO and 200μmol/BHA,and harvested 5h later;（2）The control group with the same induction method but receiving no Aβ25-35.Morphological changes were observed by light microscope,neuron-specific enolase（NSE）were detected by immunocytochemistry;GSK-3β,Tau[pSer262]and Tau[pSer396]were tested by Western blot.Results with the observation of light microscope,spindle shaped BMSCs after induction showed neuron-like cells shape with long projections,which were different from Aβ25-35 experiment group with the length and quantity of the projections shorter and less;both groups showed Tau＋by immunocytochemistry;Western blotting showed that GSK-3β,Tau[pSer262]and Tau[pSer396]of Aβ25-35 treatment group were significantly higher than those of the control group.Conclusion Aβ25-35 affected BMSCs differentiated neuron-like cells with the hyperphosphorylation of Tau protein through GSK-3.
出处
《中风与神经疾病杂志》
CAS
CSCD
北大核心
2011年第7期580-582,共3页
Journal of Apoplexy and Nervous Diseases
