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目的 研究电脉冲介导的基因转移效率及其最佳基因转移电脉冲参数。方法 用微量注射器将pcDNA3-Luc质粒100μg注射入BALB/小鼠的股四头肌,5 min内在注射部位给予不同参数的电脉冲刺激,然后进行荧光素酶活性测定和组织病理切片。结果 电脉冲可明显增加荧光素酶基因的表达,电脉冲组的荧光素酶活性(7.26±3.72)×107 RLU/mg蛋白是单纯注射组(1.38±1.27)×104 RLU/mg蛋白的5 262倍(P=0.017)。组织病理检查发现电脉冲刺激无严重的肌肉组织损伤。电压100 V/cm,波宽60 ms,脉冲次数10次和频率0.75 Hz是最佳的电脉冲参数。结论 在最佳的电脉冲参数条件下,电脉冲介导的基因转移可获得较高的基因表达。
OBJECTIVE: To study the efficiency of electrical pulse-mediated gene transfer and its optimal gene transfer electrical pulse parameters. Methods 100 μg of pcDNA3-Luc plasmid was injected into the quadriceps femoris of BALB / mouse with a microinjector and stimulated with different parameters at 5 min after injection. The luciferase activity and histopathological sections were also measured. Results The electric pulse significantly increased the luciferase gene expression. The electro-pulsed luciferase activity (7.26 ± 3.72) × 107 RLU / mg protein was 5 262 (1.38 ± 1.27) × 104 RLU / mg protein Times (P = 0.017). Histopathological examination revealed that electrical pulses stimulate no serious muscle tissue damage. Voltage 100 V / cm, wave width 60 ms, number of pulses 10 times and frequency 0.75 Hz are the best electrical pulse parameters. Conclusion Electrical pulse-mediated gene transfer can achieve higher gene expression under the optimal electrical pulse parameters.