BDNF-GFP转染神经干细胞对脊髓损伤大鼠BDNF表达的影响  

Effect of brain-derived neurotrophic factor and green fluorescent protein gene-transfected neural stem cells transplantation on brain-derived neurotrophic factor expression in rats with spinal cord injury

作　　者：王岩松[1] 梅晰凡[1] 吕刚[1] 

机构地区：[1]辽宁医学院附属第一医院骨科,辽宁锦州121000

出　　处：《军医进修学院学报》2011年第4期369-372,共4页Academic Journal of Pla Postgraduate Medical School

基　　金：辽宁省自然科学基金(20052096)~~

摘　　要：目的探讨脑源性神经营养因子(Brain-Derived Neurotrophic Factor,BDNF)和绿色荧光蛋白(Green Fluorescent Protein,GFP)转染后神经干细胞(Neural Stem Cells,NSCs)移植对脊髓损伤大鼠BDNF表达的影响。方法以携带BDNF-GFP基因的腺病毒转染NSCs,免疫组化及Western blot检测转染后NSCs BDNF表达。40只Wistar大鼠中假手术组8只,32只大鼠制成T9左侧横断模型,并随机分成四组:BDNF和GFP修饰的N-SCs移植组、GFP修饰的NSCs移植组、单纯NSCs移植组和模型组。在各NSCs移植组,脊髓损伤后向横断处显微注射等体积细胞,模型组在相同的部位注射等体积的PBS。实时定量PCR检测各实验组BDNF表达情况。结果免疫组化显示BDNF-GFP转染的NSCs可表达BDNF(黄色荧光)。Western blot结果显示BDNF-GFP转染的NSCs可表达相对分子质量约为41kU的融合蛋白条带。NSCs移植可使BDNF的表达量明显增高(P<0.05或P<0.01),以BDNF-GFP转染的NSCs移植组BDNF表达量最高(P<0.01)。结论 BDNF-GFP转染后NSCs可在脊髓半切模型中存活并高表达具有生物活性的BDNF。Objective To study the effect of brain-derived neurotrophic factor（BDNF） and green fluorescent protein（GFP）-transfected neural stem cells（NSCs） transplantation on expression of BDNF in rats with spinal cord injury.Methods NSCs were transfected with adenovirus vector bearing BDNF and GFP.Expression of BDNF in BDNF and GFP-transfected NSCs was detected by immunohistochemistry and Western blot,respectively.Of the 40 healthy Wistar rats,8 were selected as a sham-operation group,32 served as a T9 left hemisection model.Then,the 32 rats were randomly divided into BDNF and GFP-transfected NSCs transplantation group,GFP-transfected NSCs transplantation group,single NSCs transplantation group and model groups,8 rats in each group.Gene-transfected NSCs or non gene–transfected NSCs were microinjected into each side of the transection site in the 3 NSCs transplantation groups after spinal cord injury（SCI） was induced.An equal volume of PBS was injected into the model group through the same injection sites.Expression of BDNF was detected in each group after SCI by real-time PCR.Results Immunohistochemistry showed that BDNF and GFP-transfected NSCs could express BDNF（yellow fluorescence）.Western blot demonstrated that BDNF and GFP-transfected NSCs could express immunoreactive bands with a relative molecular mass of 41kU.NSCs transplantation could significantly increase the expression level of BDNF（P0.01）.The expression level of BDNF was the highest in BDNF and GFP-transfected NSCs transplantation group（P0.01）.Conclusion BDNF and GFP-transfected NSCs can survive and highly express BDNF in hemisected spinal cord model of rats.

关 键 词：神经元细胞 脑源性神经营养因子 绿色荧光蛋白质类 脊髓损伤 移植 

分 类 号：R34[医药卫生—基础医学]