脂质体介导胶质细胞源性神经营养因子基因在体骨骼肌的转移和表达

Expression of glial cell line-derived neurotrophic factor gene mediated by liposome in skeletal muscle in vivo

目的：研究脂质体介导胶质细胞源性神经营养因子(GDNF)基因在大鼠骨骼肌内的表达。方法：将鼠 GDNF cDNA 克隆到真核表达载体 pEGFP 中,构建重组载体 pEGFP-GDNF；将脂质体和重组质粒 pEGFP-GD- NF cDNA 混合后直接注入大鼠面神经支配的骨骼肌内。采用免疫组化技术和荧光显微镜检测 GDNF 基因转染后的体内表达。结果：转染2 d 后,转染侧面肌细胞内有散在的绿色荧光,且有部分细胞呈 GDNF 免疫反应阳性,对照侧为阴性。结论：GDNF 基因能通过脂质体介导转入骨骼肌内并表达 GDNF 蛋白,为神经营养因子基因肌内转染治疗周围神经损伤提供了初步的理论和实验依据。

Objective： To examine the expression of glial cell line-derived neurotrophic factor （GDNF） in skeletal muscle of rats. Methods： The recombinant plasmid pEGFP-GDNF was constructed by cloning GDNF eDNA into eukaryotic expession vector pEGFP. The complex of recombinant plasmid pEGFP-GDNF and liposome were infected into the skeletal muscles innerated by facial nerve of rats. The expressions of GDNF and GFP in the skeletal muscles were examined using immunohistochemical staining and fluorescence microscope. Results： Two days after the injection, GDNF postive staining and green fluoresence substances were found in muscle cells of the experimental side but not in control side. Conclusion： GDNF gene can be transduced into and expressed in muscle cells.