论文部分内容阅读
为了观察质粒型单纯疱疹病毒载体介导的外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因在体外培养的金黄地鼠肾细胞以及脊髓神经元和背根节神经元中的转移和表达 ,本研究采用了以质粒型单纯疱疹病毒载体为基础构建的分别含有重组胶质细胞源性神经营养因子和绿色荧光蛋白基因的混合毒株 dv HSV-GDNF和 dv HSV-GFP感染体外培养的金黄地鼠肾细胞、脊髓神经元和背根节神经元。用免疫组织化学方法和荧光显微镜观测法分别检测了胶质细胞源性神经营养因子和绿色荧光蛋白基因的转移和表达。结果发现 :质粒型单纯疱疹病毒载体可以成功地将外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因导入金黄地鼠的肾细胞以及脊髓神经元和背根节神经元中。提示质粒型单纯疱疹病毒载体可以作为转基因胶质细胞源性营养因子治疗脊髓损伤的转移载体 ,为脊髓损伤的基因治疗提供了实验基础。绿色荧光蛋白作为报告基因 ,也可因其适用广泛、观察简便等特性而得到更加广泛的应用。
In order to observe the transfer of exogenous glial cell line-derived neurotrophic factor and green fluorescent protein gene mediated by plasmid herpesvirus vector in golden hamster kidney cells and spinal cord neurons and dorsal root ganglion neurons in vitro and In this study, we used in vitro culture of mixed strains dv HSV-GDNF and dv HSV-GFP with recombinant glial cell line-derived neurotrophic factor and green fluorescent protein respectively based on plasmid herpes simplex virus vector Golden hamster kidney cells, spinal cord neurons and dorsal root ganglion neurons. Immunohistochemistry and fluorescence microscopy were used to detect the expression of glial cell line-derived neurotrophic factor (EGFR) and green fluorescent protein (EGFP) gene. The results showed that the plasmid herpes simplex virus vector successfully transfected exogenous glial derived neurotrophic factor and green fluorescent protein gene into golden hamster kidney cells as well as spinal cord neurons and dorsal root ganglion neurons. It is suggested that the plasmid herpes simplex virus vector can be used as a transfer carrier of genetically modified glia derived factor for the treatment of spinal cord injury and provide experimental basis for gene therapy of spinal cord injury. Green fluorescent protein as a reporter gene, but also because of its wide range of applications, such as observation is simple and more features and be more widely used.