超声介导声诺维提高人血管内皮细胞基因转染效率的体外实验研究

目的探讨使用超声破坏微泡造影剂-声诺维的方法对脂质体介导的增强型绿色荧光蛋白(EPGFP)基因转染人脐静脉血管内皮细胞(HUVEC)的作用。方法在单孔培养皿中加入HUVEC细胞悬液,每孔加入脂质体介导EPGFP4μg,加或不加微泡造影剂或不同条件的超声辐照。24h后,用荧光显微镜及流式细胞仪测量EPGFP在细胞内的表达及基因的转染效率。结果超声组转染率提高,超声 微泡组转染率明显提高。超声 微泡组,超声辐照时间60s机械指数(MI)1.4组转染效率最高;超声机械指数1.0辐照时间90s组转染率最高。结论声诺维在超声作用下能明显提高脂质体介导的基因对细胞转染的效率,可能作为基因治疗的载体。

High-eff iciency gene transfection of HUVEC via ultrasound-mediated destruction of mirobubbles in vitro

Objective To investigate the effects of ultrasound-mediated destruction of mirobubbles on HUVEC transfected with lipofectamine-EPGFP. Methods HUVEC suspension was in single-well plate, lipofectamine complexed with EPGFP 4ug were transfected to HUVEC, with or without ultrasound contrast agents-SonoVue and with or without treatment of different characteristics ultrasound. Expression of the marker gene EPGFP was quantified by FACS analysis and observed by fluorescent after 24 hours. Results US treatment of HUVEC on plates yielded a increase transfection rates, US-mediated destruction of SonoVue of HUVEC on plates yielded a significant increase transfection rates. US treatment of MI 1.4 for 60 s led to the highest transfection effect; US treatment for 90 s under MI 1.0 led to the highest transfection effect. Conclusion Ultrasound-mediated destruction of SonoVue mirobubbles enhances transfection rates of gene after lipofectamine-mediated to HUVEC in vitro. SonoVue can be used as vectors in gene therapy.