中国组织工程研究2012，Vol.16Issue(15)：2704-2708,5.DOI:10.3969/j.issn.1673-8225.2012.15.010
六氟化硫微泡介导人脂联素基因兔主动脉内转染及表达
Transfection and expression of human apM1 gene into the aorta of rabbits mediated by using ultrasound with SonoVue
摘要
Abstract
BACKGROUND: Gene delivery using ultrasound microbubbles is an emerging directional transmission gene technology. OBJECTIVE: To construct a recombinant plasmid containing human apM1 gene and delivery apM1 gene into the aorta of rabbits by using ultrasound with SonoVue. METHODS: Twenty-one rabbits were randomized into control group (n=6) and apM1 group (n=15). apM1 mRNA from epicardial adipose tissue was amplified by reverse transcription-PCR and cloned into plasmid pcDNA3.1 to generate the recombinant plasmid pcDNA3.1-apM1, which was confirmed by nucleotide sequencing and transiently transfected into human umbilical vein endothelial cells (HUVECs). While the mixture of SonoVue and pcDNA3.1-apM1 was injected though helix vein, the thoracoabdominal aorta of rabbits were sonicated by ultrasound for 3 minutes. The arterial vessels and blood serum of rabbits were harvested at 2, 7and 14 days respectively after delivery to detect apM1 expression by Western blot analyses and enzyme linked immunosorbent assay (ELISA). RESULTS AND CONCLUSION: The identification of pcDNA3.1-apM1 recombinant by DNA sequencing ws correct. The expression of apM1 gene could be detected by Western blot in HUVECs after pcDNA3.1-apM1 transient transfection. apM1 gene could be effectively transferred into the aorta wall of rabbit by using ultrasound with SonoVue. The high expression of apMl gene was detected at 2 days after delivery and until to 14 days. ELISA results showed that the concentration of apM1 protein in serum of rabbit could be significantly increased by delivery of apM1 gene using ultrasound with SonoVue (P < 0.01). These findings indicate that the eukaryotic expression recombinant containing apM1 gene can be successfully constructed. apM1 gene can be transferred into the deep area of rabbit aorta vessel wall by ultrasound with SonoVue and expressed and secreted effectively.关键词
六氟化硫微泡/脂联素/动脉粥样硬化/基因/重组质粒分类
医药卫生引用本文复制引用
杨文凯,陈庆,杨春丽,王现国,谢霆,董念国,陈新忠..六氟化硫微泡介导人脂联素基因兔主动脉内转染及表达[J].中国组织工程研究,2012,16(15):2704-2708,5.基金项目
国家自然科学基金项目(30872541) （30872541）
博士点基金新教师项目资助课题(200804871116). （200804871116）
