【摘要】 目的:探讨超声介导SonoVue微泡破裂法促进外源基因在肝脏中表达的作用,评估其转染效率、安全性。方法:经SD大鼠门静脉置管输入SonoVue微泡建立动物模型,以虫荧光素酶基因为报告基因,观察不同浓度的微泡促进虫荧光素酶基因在肝脏中的表达强度与持续时间,实验结束后采下腔静脉血检测肝肾功能并取肝、肾组织常规HE染色行组织学检查。结果:随着微泡浓度的增加,虫荧光素酶基因表达逐渐增加,在微泡浓度为100%(V/V)时,转染效率最高,持续到21天仍有表达。转染前后的肝肾功能(AST、ALT、ALB、BUN、Cr)无明显变化,HE切片未发现肝肾组织结构的改变。结论:超声介导微泡破裂法能有效地促进外源基因在肝脏中的表达,为肝脏疾病的基因治疗提供了新的途径。更多还原

【Abstract】 Objective: To evaluate the efficiency and safety of gene transfer by ultrasound mediated microbubbles destruction in liver in vivo. Methods:Plasmid pGL3 containing Luciferase gene, mixtures of different concentration SonoVue microbubbles and plasmid were transfected into the rat liver via portal vein under ultrasound exposure to test the optimized concentration of microbubbles. Luciferase activity was measured on the 4th, 7th, 15th and 21st day after gene transfer respectively. The hepatic and renal function were monitored during experiments, and the histological study of liver and kidney was performed after experiments to evaluate the potential effects of gene transfer. Results: Ultrasound mediated microbubbles destruction significantly enhance the efficiency of target gene transfer into the liver. A dose-dependent effect of microbubbles was observed SonoVue5%:(55 ± 10) RLU/(mg·prot·min); SonoVue50%:(609 ± 82)RLU/(mg·prot·min); SonoVue100%:(827 ± 124) RLU/(mg·prot·min), P < 0.01. The serum biochemical and histological analyses did not reveal harmful effect. Conclusion: Ultrasound mediated microbubble destruction method enhanced the efficiency of Luciferase gene transfer into the liver in rats. This non-viral method might become a new approach to gene therapy for hepatic disease.更多还原