脑源性神经营养因子过表达促进大鼠神经干细胞向神经元分化  

Brain-derived neurotrophic factor overexpression promoting the differentiation of rat neural stem cells into neurons

作　　者：欧阳长杰[1] 滕大才[1] 曲德伟[1] 王德广[1] 徐铁军[1] 

机构地区：[1]徐州医学院人体解剖学教研室,江苏徐州221002

出　　处：《解剖学报》2011年第6期726-730,共5页Acta Anatomica Sinica

基　　金：江苏省自然科学基金资助项目(BK2009087);徐州医学院科研资助项目(2010KJ17);徐州市科技计划资助项目(XM09B084)

摘　　要：目的构建脑源性神经营养因子(BDNF)基因真核表达载体,探讨BDNF过表达对神经干细胞(NSCs)向神经元分化的影响。方法采用RT-PCR法,以大鼠海马组织RNA为模板,扩增BDNF基因,定向克隆到pEGFP-N1载体中,用脂质体法转染pEGFP-N1-BDNF表达载体至NSCs中,然后用RT-PCR鉴定BDNF的表达,免疫组织化学方法鉴定NSCs向神经元的分化情况。结果成功构建了pEGFP-N1-BDNF真核表达载体,BDNF在重组质粒转染的NSCs中能够高效表达。重组质粒转染的NSCs在体外诱导分化后,能够较空质粒转染的NSCs产生更多的神经元(P<0.01)。结论 BDNF过表达能够显著促进大鼠NSCs向神经元方向分化。Objective To construct eukaryotic expression vector of brain-derived neurotrophic factor （BDNF） and detect its effect of overexpression on differentiation of rat neural stem cells （NSCs） into neurons. Methods The RT-PCR was used to amplify rat BDNF gene from RNA of rat hippocampus. The BDNF gene was inserted into eukaryotic expression vector pEGFP-N1 to construct recombinant expression vector pEGFP-N1-BDNF. The recombinant vector was transfected into NSCs by Lipofectamine 2000. The expression of BDNF mRNA in NSCs was detected by RT-PCR. The differentiation of rat NSCs into neurons was detected by immunohistochemistry staining. Results The sequence of the cloned BDNF was confirmed to be correct by DNA sequencing. The NSCs transfected with pEGFP-N1-BDNF expressed BDNF efficiently. The pEGFP-N1-BDNF transfected NSCs differentiated into more neurons than the pEGFP-N1 transfected ones （ P 〈 0. 01 ）. Conclusion All these results indicate that BDNF overexpression significantly promotes the differentiation of rat NSCs into neurons.

关 键 词：脑源性神经营养因子 基因表达 神经干细胞 分化 反转录-聚合酶链式反应 免疫组织化学 大鼠 

分 类 号：Q786[生物学—分子生物学]