逆转录病毒介导的脑源性神经营养因子在大鼠成肌细胞中表达及分泌  

Brain derived neurotrophic factor expression and secretion in rat myoblast genetically modified with a retroviral vector

作　　者：曲伸[1] 何晓龙[1] 王成海[1] 路长林[1] 

机构地区：[1]第二军医大学基础医学部神经生物学教研室

出　　处：《第二军医大学学报》1997年第3期219-221,共3页Academic Journal of Second Military Medical University

摘　　要：目的：研究经逆转录病毒载体的介导将脑源性神经营养因子（ＢＤＮＦ）ｃＤＮＡ导入大鼠成肌细胞，为体内移植治疗打下基础。方法：采用电穿孔法将重组大鼠ＢＤＮＦｃＤＮＡ导入ＰＡ３１７包装细胞，获取高滴度的病毒上清转染成肌细胞。Ｇ４１８筛选转染的成肌细胞，采用Ｎｏｒｔｈｅｍ杂交和ＰＣ１２细胞存活检测。结果：证实转染细胞内有ＢＤＮＦｍＲＮＡ的表达并具有促进细胞存活的生物学活性。结论：ＢＤＮＦｃＤＮＡ可在真核细胞内表达并具有生物学效应，可用于神经系统基因治疗的研究。Objective:To introduce brain derived neurotrophic factor(BDNF)cDNA (BDNF cDNA) into rat myoblast with a retroviral vector PN 2A for the treatment of CNS disease in vivo.Methods:Recombined BDNF cDNA plasmid was transfected by the electroporation into the packing cell line PA317 and G418 resistant clones with highest titer was selected.Myoblastas L 6TG were infected with virus supernatant and selected under G418,then transgenic BDNF mRNA synthesis was screened by Northern blotting.Results:Northern blot revealed that there were BDNF mRNA expressions in L 6TG cells and in vitro bioassay showed that the conditioned medium of engineered myoblasts could promote the neuritis outgrowth of PC12 and the concentration of BDNF had the linear correlation with the survival of the PC12 cells.Conclusion:It suggests that these cells genetically modified can express BDNF in vitro and may be used as a tool of intracranial gene therapy of CNS disease.

关 键 词：逆转录病毒 神经营养因子 脑源性 成肌细胞 

分 类 号：R338.1[医药卫生—人体生理学] Q753[医药卫生—基础医学]