脂质超声微泡增强AAV介导的致密结构细胞的基因转导  

Lipid ultrasound microbubble augmentated AAV-mediated gene transduction in cells with tight junction

作　　者：刘中兵[1] 钟志容[2] 傅秀娟[2] 李劲薇[2] 兰欢[3] 余昕[2] 何颖[2] 左正云[2] 

机构地区：[1]泸州医学院现代教育技术中心,四川泸州646000 [2]泸州医学院药学院,四川泸州646000 [3]泸州医学院心血管医学研究所,四川泸州646000

出　　处：《泸州医学院学报》2013年第1期1-5,共5页Journal of Luzhou Medical College

基　　金：国家自然科学基金项目(81202479);教育部重点项目(212148);四川省教育厅重点项目(10ZA039);四川省卫生厅项目(100225)

摘　　要：目的:探讨超声介导的脂质微泡能否增强腺相关病毒(AAV)对具有致密结构的细胞的转导效率。方法:制备载AAV-eGFP的脂质超声微泡,以Malvern粒度检测仪检测微泡粒径及分布,选用人支气管上皮细胞16HBE14o-为模型细胞,采用Transwell小室细胞培养技术培养具有紧密结构的人支气管上皮细胞16HBE14o-,利用倒置荧光显微镜定性观察转导效率,采用流式细胞技术进一步定量检测转导效率。结果:本试验制备的载AAV脂质超声微泡呈圆球形均匀分布,平均粒径为2.46±1.2μm;脂质微泡携同超声处理能显著增强AAV对形成致密结构的人支气管上皮细胞16HBE14o-转导效率。结论:超声介导脂质微泡为AAV在基因治疗中的应用提供了一种新的途径,为AAV在体内的靶向应用奠定了基础。Objective： To test whether the lipid microbubble exposed to ultrasound could enhance the adenoassociated virus （AAV） mediated gene transduction in cells with tight junction. Methods： The AAV-loaded lipid ultrasound microbubble was prepared by the bath-ultrasound method and the particle size or distribution was measured by Malvern Sizer Nano ZS90. The human bronchial epithelial cells of 16HBE14o- were selected as the model cells, which were cultured in the Transwell insert to form the tight junction between cells of 16HBE14o-. After administration, the transducted cells were observed for positive cells with expression of green fluorescence protein. Further, the transduction efficiency was estimated by the flow cytometric analysis. Results： The AAV- loaded lipid ultrasound microbubble prepared in this experiment was spherical with uniform distribution and with an average particle size of 2.46±1.2 μm. It could significantly enhance the gene transduction of 16HBE14o- cells after ultrasound exposure. Conclusion： The lipid ultrasound microbubble medicated by ultrasound generated a novel strategy for AAV in the gene therapy application, which supplies the basis for AAV target using in viva.

关 键 词：脂质超声微泡 腺相关病毒 基因转导 人支气管上皮细胞 

分 类 号：Q54[生物学—生物化学]