Electroporation-facilitated delivery of plasmid DNA in skeletal muscle: Plasmid dependence of muscle damage and effect of poloxamer 188
Electroporation-facilitated delivery of plasmid DNA in skeletal muscle: Plasmid dependence of muscle damage and effect of poloxamer 188
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DOI：
10.1006/mthe.2001.0483
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发表时间：
2001-11-01
影响因子：
12.4
通讯作者：
Sawdey, M
中科院分区：
文献类型：
--
作者：
Hartikka, J;Sukhu, I;Sawdey, M
据报道，电穿孔可以促进骨骼肌中的裸DNA基因转移，但也与电损伤的发病机制有关。为了评估电穿孔对基因转移的影响，将荧光素酶报告质粒VR1255注射到小鼠四头肌肌肉中，并用卡尺电极电穿孔。肌内荧光素酶的表达通过电穿孔增加10- 70倍，这取决于所使用的DNA剂量和注射量。在没有质粒DNA注射的情况下，股四头肌电穿孔导致血清肌酸磷酸激酶活性迅速升高，但没有引起明显的肌肉损伤。然而，在注射质粒DNA并电穿孔的肌肉中，当施加最适合基因转移的电场强度（300伏/厘米）时，在电极放置的近端区域持续出现可见病变。肌肉病变的发展与质粒转基因表达无关，在电穿孔过程中需要质粒在肌肉中存在。poloxam188 （pluronic F68）与VR1255联合注射可显著降低电穿孔后血清肌酸磷酸激酶活性的升高，但不能抑制肌肉病变的发展。在非电穿孔肌肉中，联合注射poloxam188使荧光素酶的表达增加了三倍。因此，波洛沙姆188可构成肌内递送裸DNA的有用赋形剂。
Electroporation has been reported to facilitate naked DNA gene transfer in skeletal muscle, but has also been implicated in the pathogenesis of electrical injuries. To assess the effects of electroporation on gene transfer, mouse quadriceps muscles were injected with the luciferase reporter plasmid VR1255 and electroporated with caliper electrodes. Intramuscular luciferase expression was increased 10- to 70-fold by electroporation, depending on the DNA dose and injection volume used. In the absence of plasmid DNA injection, electroporation of quadriceps muscles resulted in rapid elevations in serum creatine phosphokinase activity, but did not elicit visible muscle damage. However, in muscles injected with plasmid DNA and electroporated, visible lesions consistently developed in the areas proximal to electrode placement when field strengths optimal for gene transfer (300 volts/cm) were applied. The development of muscle lesions was independent of plasmid transgene expression and required the presence of plasmid in the muscle during electroporation. Co-injection of poloxamer 188 (pluronic F68) with VR1255 substantially reduced elevations in serum creatine phosphokinase activity following electroporation, but did not inhibit the development of muscle lesions. In non-electroporated muscles, co-injection of poloxamer 188 increased luciferase expression threefold. Poloxamer 188 may thus constitute a useful excipient for intramuscular delivery of naked DNA.