神经型钙黏蛋白对小鼠脂肪间充质干细胞的成神经作用  

Role of N-cadherin in inducing adipose tissue-derived mesenchymal stem cells into nerve cells

作　　者：贾阳阳[1] 杨慈清[1] 付苏雷[1] 李晗[1] 郭志坤[1] 骆建凯 

机构地区：[1]新乡医学院河南省医用组织再生重点实验室

出　　处：《解剖学杂志》2015年第6期659-659,671,封4,共6页Chinese Journal of Anatomy

摘　　要：目的:探讨神经型钙黏蛋白(N-cadherin)在小鼠脂肪间充质干细胞(ADMSCs)成神经分化中的作用.方法;用差速培养法获取小鼠ADMSCs,传至第5代,经成神经诱导液诱导ADMSCs 7 d后,免疫荧光检测胶质纤维酸性蛋白(GFAP)、神经元特异性烯醇化酶(NSE)和微管相关蛋白2(MAP2)的表达,半定量PCR检测细胞中N-cadherinmRNA的表达;采用N-cadherin对细胞进行基因修饰,免疫荧光检测神经丝蛋白(NF)和GFAP的表达.结果:ADMSCs经成神经诱导7d后表达GFAP、NSE和MAP2,半定量PCR结果显示,与对照组相比成神经诱导后的细胞表达N-cadherin显著增高.转染N-cadherin的N-cadherin细胞培养24h后发出细长的突起,与相邻细胞之间形成网状连接.免疫荧光结果显示,转染后的细胞NF和GFAP表达阳性.结论:ADMSCs在体外多种作用下,具有向神经细胞分化的潜能.N-cadherin转染后能改变ADMSCs的形态,并且在ADMSCs向神经分化中发挥一定的作用.Objective:To explore the function of N-cadherin in inducing adipose tissue-derived mesenchymal stem cells(ADMSCs) isolated from mice into nerve cells.Methods:ADMSCs were isolated from mice by differential culture method and the fifth generation ADMSCs were used for this study.Immunofluorescent method was used to detect the expression of glial fibrillary acidic protein(GFAP),neuron specific enolization enzyme(NSE) and microtubule associated protein 2(MAP2)after being induced for 7 days;The expression of N-cadherin in ADMSCs was detected by RT-PCR.And then ADMSCs were transfected with N-cadherin and evaluated the expression of neurofilament protein(NF),GFAP by immunofluorescent method.Results:ADMSCs could express GFAP,NSE and MAP2 after being induced for 7 days.The expression of Ncadherin mRNA was higher than the control group by RT-PCR.Immunofluorescent images showed that ADMSCs could send out long protrusions to form network connections and could express NF and GFAP after being transfected with N-cadherin for24 hours.Conclusion:ADMSCs isolated from mice have nerve differentiation potential in vitro.N-cadherin can change the morphology of ADMSCs significantly,and plays an important role in prompting ADMSCs to differentiate into astrocytes and neurons.

关 键 词：神经型钙黏蛋白 脂肪间充质干细胞 成神经诱导 半定量PCR 转染 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]