The distribution and time course of brain-derived neurotrophic factor expression after plasmid DNA transfer to adult rat spinal cord

我们研究了非病毒载体介导的脑源性神经营养因子（BDNF）基因转染成年大鼠脊髓的潜力。使用三种阳离子脂质（lipofecectin ^<TM>, LipofectAMINE^<TM>和LipofectAMINE^<TM> 2000）检测转染体内脊髓的潜力，这三种脂质已被证明可以在体外使用质粒DNA将基因转染到神经细胞中。将脂质体cDNA复合物直接注入胸脊髓后，组织学分析和免疫印迹分析显示，只有LipofectAMINE^<TM> 2000/ cDNA复合物组有绿色荧光蛋白（GFP）和外源性BDNF的表达。在LipofectAMINE^<TM> 2000/ cDNA复合物组中，荧光组织化学检查证实注射后至少5天注射区周围存在注射GFP。转染后的细胞被鉴定为反应性星形胶质细胞和部分神经元。RT-PCR检测的BDNF mRNA和Western Blotting检测的BDNF蛋白均显示出相似的特征。结果表明，阳离子脂质体介导的传递可能是一种实用的脊髓基因转移方法。

We investigated the potential of non-viral vector-mediated brain-derived neurotrophic factor (BDNF) gene transfection in adult rat spinal cord. The potential of transfection of in vivo spinal cord was examined using three cationic lipids, Lipofectin^<TM>, LipofectAMINE^<TM> and LipofectAMINE^<TM> 2000, which have been proven to transfect genes into neuronal cells in vitro using plasmid DNA. After the direct injection of liposome cDNA complexes into thoracic spinal cord, histological analysis and immunoblot analysis demonstrated green fluorescent protein (GFP) and exogenous BDNF expression only in the group of LipofectAMINE^<TM> 2000/ cDNA complexes. In the group of LipofectAMINE^<TM> 2000/ cDNA complexes, fluorescent histochemical examination confirmed the presence of injected GFP around the injection areas for at least 5 days after injection. The transfected cells were identified reactive astrocytes, and some neuron. Both BDNF mRNA using RT-PCR and BDNF proteins using Western Blotting indicated a similar profile. The results suggest that cationic liposome-mediated delivery may be a practical method for gene transfer in spinal cord.