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目的:探究电激和普鲁卡因对DNA导入哺乳动物活体效率的影响。方法:向小鼠的股四头肌注射20μgβ-gal质粒,分别采用以下方法:直接注射;先注射100μL 0.25%的普鲁卡因,24 h后注射质粒;注射质粒后进行经过前期优化后的电激程序进行加强(500 V,4个脉冲)。注射后的第2 d、1w和2w后进行x-gal染色检测。结果:三种导入方法均可将外源质粒导入肌肉组织中并进行表达。电激和普鲁卡因可以显著增强肌肉对外源基因的吸收和表达,尤其是优化后的电激程序,其可以使肌肉长期稳定高效的表达外源导入基因。结论:普鲁卡因是常用的增强DNA导入效率的促进剂,而合适的电激可以更为显著地增强DNA导入效率,均是基因治疗中提高基因导入效率的良好候选方案。
Objective: To investigate the effects of electroporation and procaine on the in vivo efficiency of DNA into mammals. Methods: 20μg β-gal plasmids were injected into the quadriceps femoris of rats. The mice were injected directly with 100μL of 0.25% procaine and injected with plasmids after 24 h. Plasmid was injected and pre-optimized Electro-excitation procedure is enhanced (500 V, 4 pulses). After 2 d, 1 w and 2 w after injection, x-gal staining was performed. Results: Three kinds of introduction methods can be used to import foreign plasmid into muscle tissue and express it. Electrical stimulation and procaine can significantly enhance muscle absorption and expression of foreign genes, especially the optimized electroporation program, which can make the muscle stably and efficiently express exogenous genes for a long time. Conclusion: Procaine is a commonly used promoter to enhance the efficiency of DNA introduction. Appropriate electroporation can enhance the efficiency of DNA introduction more significantly, which is a good candidate for gene transfection in gene therapy.