S100A4促进脑源性神经营养因子表达影响神经干细胞的分化  

S100A4 promotes differentiation of neural stem cells through up-regulation of brain-derived neurotrophic factor

作　　者：杜晓文 林大鹏 屠冠军[1] Du Xiaowen;Lin Dapeng;Tu Guanjun(Department of Orthopedics,the First Affiliated Hospital of China Medical University,Shenyang 110001,Liaoning Province,China;Department of Orthopedics,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China)

机构地区：[1]中国医科大学附属第一医院骨科,辽宁省沈阳市110001 [2]锦州医科大学附属第一医院骨科,辽宁省锦州市121001

出　　处：《中国组织工程研究》2020年第19期3029-3034,共6页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省自然科学基金指导计划项目(201602857)，项目负责人：屠冠军~~

摘　　要：背景:如何促进神经干细胞大量向神经元分化是研究的难点。研究发现,S100A4蛋白可能通过多种途径在中枢神经系统修复中发挥作用。目的:研究S100A4是否通过上调脑源性神经营养因子表达进而影响神经干细胞向神经元样细胞的分化。方法:购买鉴定合格的小鼠胚胎大脑海马和室管膜下区来源的神经干细胞,体外培养传代,电穿孔法向神经干细胞中转染S100A4表达载体和/或脑源性神经营养因子siRNA,转染48 h后向神经元方向诱导分化,诱导分化3 d后采用Western blot检测细胞中脑源性神经营养因子及Tuj1蛋白表达,免疫荧光检测Tuj1阳性神经元比例。结果与结论:①与转染无关序列质粒组比较,转染S100A4组神经干细胞中Tuj1阳性细胞比例及相应的Tuj1、脑源性神经营养因子的表达量显著增高(P<0.01);②与S100A4+siRNA无关序列共转染组比较,S100A4+脑源性神经营养因子siRNA共转染组神经干细胞中Tuj1阳性细胞比例及相应的Tuj1、脑源性神经生长因子的表达量显著降低(P<0.01);③结果表明,S100A4的过表达可促进神经干细胞向神经元样细胞分化,其可能通过脑源性神经营养因子发挥作用。BACKGROUND:How to promote neural stem cells differentiate into neurons is a difficulty.S100A4 has been found to play a role in the nervous system repair by various pathways.OBJECTIVE:To investigate whether S100A4 affects the differentiation of neural stem cells into neurons through up-regulating the expression of brain-derived neurotrophic facto.METHODS:The neural stem cells from brain hippocampus and subependymal region of embryonic mice were cultured in vitro and passaged.The S100A4 expression vector and/or brain-derived neurotrophic factor+siRNA were transfected into neural stem cells by electroporation,and the cells were induced to differentiate into neurons at 48 hours after transfection.Three days later,the expression levels of brain-derived neurotrophic factor and Tuj1 in cells were detected by western blot assay.Proportion of Tuj1 positive neurons was tested by immunofluorescence.RESULTS AND CONCLUSION:Compared with the unrelated sequence plasmid group,the proportion of Tuj1 positive neurons and the expression levels of Tuj1 and brain-derived neurotrophic factor in the S100A4 transfection group were significantly increased(P<0.01).Compared with the S100A4+siRNA unrelated sequence plasmid group,the proportion of Tuj1 positive neurons and the expression levels of Tuj1 and brain-derived neurotrophic factor in the co-transfection group were significantly decreased(P<0.01).These results indicate that S100A4 overexpression can promote the differentiation of neural stem cells into neurons,which may be mediated by brain-derived neurotrophic factor.

关 键 词：脊髓损伤 S100A4 神经干细胞 电穿孔 神经元 细胞分化 脑源性神经营养因子 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]