中文摘要：

目的以遗传性视网膜色素变性RCS大鼠为模型，探讨在体电穿孔辅助碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）基因转染视网膜神经节细胞（retinal ganglion cell，RGC）层治疗视网膜色素变性（retinitis pigmentosa，RP）的可行性。方法将编码bFGF的质粒注射至RCS大鼠右眼玻璃体腔内，左眼注射TE buffer作为对照，随即以在体电穿孔法辅助基因转染。于1周后采用Western blot、逆转录聚合酶链反应（reverse transcription polymerase chain reaction，RT-PCR）半定量观察bFGF的表达，术后1、2、3、4周于光学显微镜下观察视网膜外核层的变化，术后1周用末端脱氧核苷酸转移酶介导的脱氧三磷酸尿苷缺口末端标记法（terminal deoxynucleotidyl transferasemediated dUTP nick end labeling，TUNEL）检测细胞凋亡情况。结果术后1、2、3、4周基因治疗眼视网膜外核层较同期对照眼厚度增加，存活的光感受器数量较对照眼多（P〈0．05）。Western blot、RT—PCR可见bFGF蛋白和mRNA的表达。TUNEL结果显示对照眼和治疗眼外核层阳性细胞凋亡指数分别为33．58％±3．72％、12．42％±2．83％，治疗眼TUNEL阳性细胞数较对照眼明显减少（P〈0．05）。结论在体电穿孔辅助bFGF基因转染RGC层能够延缓RCS大鼠视网膜色素变性的病程。

英文摘要：

Objective To investigate the feasibility of transfecting basic fibroblast growth factor （bFGF） gene into retinal ganglion cells in vivo by electroporation for preventing photoreceptor degeneration in Royal College of Sargeons （RCS） rats. Methods After intravitreous injection of the bFGF-pEGFP-N1 plasmid or TE buffer, certain square-wave electric pulses were applied to the operated eye of RCS rats. The expression of bFGF was detected by RT-PCR and Westem immunoblot analysis. The operated eyes were enucleated for histopathological examinations after 1,2,3,4 weeks. Terminal deoxynueleotidyl transferasemediated dUTP nick end labeling （TUNEL） method was used to detect the ratio of cell apoptosis in retina 1 week later. Results Eyes with injection of bFGF gene followed by in vivo electroporation showed a significantly higher level of bFGF expression in retina, a higher rescue ratio, and a lower number of TUNEL-positive photoreceptors than the control＇s （P 〈0.05）. The apoptosis index was 33.58% ± 3.72% in the control group while it was 12.42% ±2.83% in the treatment group. Conclusions Transfecting bFGF gene into retinal ganglion cells by in vivo electroporation could rescue photoreceptors in RCS rat.