论文部分内容阅读
目的:通过肌肉直接注射质粒DNA,了解局部骨骼肌细胞能否表达外源基因,并观察注射体积及注射剂量对表达的影响.方法:以β-半乳糖苷酶基因的真核细胞表达载体质粒DNA作为报告基因,通过直接肌注法观察质粒DNA在新西兰兔后肢骨骼肌表达情况,测定不同注射体积组及剂量组的表达效率.结果:外源基因能够在骨骼肌细胞内表达,表达效率随注射剂量的增加而增加,在本研究10μg、25μg、50μg、80μg和 100μg剂量组中,100μg组表达效率最高(10.30%±10.42%);注射体积对表达效率无影响,但能降低表达的变异程度.结论:基因直接肌注法能成功地转移目的基因,增加注射剂量和体积有利于提高表达效率和降低变异,该方法有望成为一种新的基因转移技术以治疗慢性下肢动脉缺血性疾病.
OBJECTIVE: To investigate whether exogenous gene can be expressed in local skeletal muscle cells by direct injection of plasmid DNA into muscle and to observe the effects of injection volume and injection dose on the expression of plasmid.Methods: The eukaryotic expression vector plasmid of β-galactosidase gene DNA as a reporter gene to observe the expression of plasmid DNA in the hindlimb skeletal muscle of New Zealand rabbits by direct intramuscular injection.Express the gene expression efficiency in different injection volume group and dose group.Results: The exogenous gene can be expressed in skeletal muscle cells with the efficiency The highest efficiency was achieved in 10μg, 25μg, 50μg, 80μg and 100μg dose groups (10.30% ± 10.42%). The injected volume had no effect on the expression efficiency but decreased the expression of the mutation .Conclusion: The direct gene transfer method can successfully transfer the target gene, increasing the dose and volume of injection can improve the expression efficiency and reduce the variation. This method is expected to become a new gene transfer technology to treat chronic lower extremity arterial ischemic disease .