超声微泡介导反义寡核苷酸转染肾脏的实验研究（英文）  

A study on the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles

作　　者：李惠玲[1] 陈金文[2] 许玄 杨儒好 向旭东[2] 张东山[2] 

机构地区：[1]中南大学湘雅二医院眼科,长沙410011 [2]中南大学湘雅二医院急诊科,长沙410011

出　　处：《中南大学学报（医学版）》2016年第2期113-120,共8页Journal of Central South University ：Medical Science

基　　金：supported by the National Natural Science Foundation(81570646);Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry(Z201508250820001);Science and Technology Project of Hunan Province(2015JC3014),P.R.China

摘　　要：目的：探讨超声联合自制脂质体微泡（lipid contrast agent,LCA）转染肾脏的条件和效率。方法：HE染色、肝肾功能检测评价其安全性;荧光显微镜评价其转染效率;RT-PCR和Western印迹检测其对结缔组织生长因子（connective tissue growth factor,CTGF）表达的影响。结果：超声联合LCA显著增强基因在肾脏内的转导效率,CY5标记的寡核苷酸主要表达在85%~90%的肾小球、肾小管和间质区;血液生化指标无明显改变;转染后第7天的LDH水平均低于0天（P〈0.05）;转染至肾脏的CTGF反义寡核苷酸成功抑制了CTGF的表达。结论：超声联合LCA的基因转移系统显著增强基因转移和表达的效率,该方法可以用来转导其他外源性的基因。Objective： To study the safety and efficiency of the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles, and to evaluate its potential clinical application.Methods： The potential and conditions regarding the transfection self-made lipid microbubbles（CY5）-labeled-oligonucleotide（ODN） or CY5-labeled-ODN connective tissue growth factor（CTGF） into the rat kidney were evaluated. The safety was evaluated by HE staining, liver and renal function tests. The transfection efficiency was evaluated by fluorescence microscopy. The expression of CTGF was detected by RT-PCR and Western blot Results： Self-made lipid microbubble and/or ultrasound significantly enhanced the efficiency of gene transfer and expression in the kidney. Especially, 85%–90% of total glomerular could be transfected. CY5-labeled-ODN expression could be observed in glomerular, tubular and interstitial area. There was no significant change in blood tests aft er gene transfer. Levels of LDH in 7 days were decreased compared with that at the first day aft er the transfection（P0.05）. CTGF expression was successfully suppressed by transfection of CTGF-antisense-ODN into kidney.Conclusion： The ultrasound-mediated gene transfer by self-made lipid microbubble could enhance the efficiency of ODN and expression in the rat kidney. Th is self-made lipid microbubbles supplement may be use for transfection of target genes.

关 键 词：超声 自制脂质微泡超声 基因治疗 

分 类 号：R450[医药卫生—治疗学]