摘要
目的 观察阳离子脂质体转染脑源性神经营养因子 (BDNF)基因对神经组织的保护和促进髓鞘再生作用。 方法 体外培养大鼠雪旺细胞 ,用阳离子脂质体介导人真核细胞质粒(PCDNA3) -BDNF重组体转移到雪旺细胞。然后将其移植到大鼠脑干网状结构损伤区 (Ⅰ组 ) ,同时设置单纯细胞移植组 (Ⅱ组 )、转染空载体细胞移植组 (Ⅲ组 )和注射等渗盐水组 (Ⅳ组 )。通过酶联免疫吸附试验检测脑内BDNF水平 ,免疫组化方法检测脑干内髓鞘碱性蛋白 (MBP) ,电镜观察髓鞘再生情况。 结果 移植后 1周 ,Ⅰ组的BDNF水平明显高于其他组 (P <0 .0 5 ) ,在免疫组化切片上可以见到Ⅰ组的MBP表达比其他组明显增多 ,透射电镜显示Ⅰ组髓鞘数量比其他组明显增多(P <0 .0 5 )。 结论 转染BDNF基因的雪旺细胞移植于损伤的脑干网状结构 ,可能有保护神经组织。
Objective To investigate the effect of brain derived neurotrophin factor (BDNF) transfected with cationic liposome in protecting nerve tissue and promoting regeneration of the myelin sheath. Methods Wistar rat Schwanns cells (SWCs) were cultured in vitro. The recombinant cationic liposomes mediated PCDNA 3 BDNF was transplanted into SWCs and then into electrical reticular lesion of the brain stem (Group Ⅰ). At the same time, pure cell transplantation group (Group Ⅱ), empty transfected vector cell transplantation (Group Ⅲ) and normal saline injection group (Group Ⅳ) were set. The concentration of BDNF in the brain stem was measured by ELISA, the survival of myelinated basic protein (MBP) and SWCs in the brain stem by immunohistochemistry and the regeneration of the myelin sheath by the transmission electron microscope (TEM). Results One week after transplantation, the concentration of BDNF in the brain stem in the Group Ⅰ was much higher than that in other three groups ( P <0.05). Two weeks after transplantation, there could be seen more MBP on the immunohistochemical slices in the GroupⅠ than that in other groups. Four weeks after the transplantation, TEM showed more significantly increased number of the myelin sheath in the Group Ⅰ compared with that in other groups ( P <0.05). Conclusions The transplantation of SWCs transfected by BDNF gene in vitro into the electrical reticular lesion of the brain stem may play a role in protecting nerve tissues and promoting the regeneration of the myelin sheath.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2003年第10期588-591,共4页
Chinese Journal of Trauma
基金
国家自然科学基金资助项目 (3 9970 75 0 )