电穿孔介导质粒基因在皮肤和线性伤口中的表达

目的 研究在线性伤口中进行电穿孔基因转染的可行性.方法 于12只Sprague-Dawley大鼠背部皮肤内注射增强型绿色荧光蛋白(EGFP)质粒(1 μg/μl)后,在注射局部进行电穿孔(800 V/cm,6个方波,每个持续20ms,间隔为200ms).电穿孔后24 h在电穿孔部位做线性切口,于第2、4、6、14天收集标本.观察EGFP表达并计算其真皮内积分光密度(IOD).然后进行HE染色.结果 电穿孔使EGFP在表皮、真皮以及肉膜内表达,真皮内表达量在第2天最高(IOD=3.50±1.45),至第14天消失,而单纯质粒注射真皮内无表达(IOD=0).结论 电穿孔能够介导质粒在线性伤口中高效而广泛地表达.

Experimental study of electroporation-mediated plasmid gene expression in skin and incisional wound

Objective To explore the feasibility of electroporation mediated gone transfer in rat incisional wound.Methods 12 Sprague-Dawley rat's dorsal skins were electroporated(800 voltages in amplitude with 6 square wsve pulses,each lasting 20 milliseconds with 200 millisecond interval)after injection of plasmid DNA(1 μg/μl,in 100 μl PBS)containing enhanced green fluorescence protein(EGFP)gene.Electroporated skins were incisiomdly wounded 24 hours after electroporation.Specimens were harvested at day 2,4,6,14,then EGFP expression in dennis wag observed and quantitatively analyzed with integrated optical density(IOD)followed by H&E staimng.Results Eleetroporation can mediate EGFP expression in epidermis,dermis and pannicuhs muscle.The expression level in dermis Wag the highest at dav 2(IOD=3.50±1.45)and disappeared at day 14.EGFP expression Wag not found in dennis if no electroporation apphed after plasmid injection(IOD=0).Conclusion Electroporation can mediate plagmid gone expression in ineisional wound efficiently and widely.