【作者】 黄强；

【导师】 冯耀良； 杨俊伟；

【作者基本信息】 南京医科大学 ， 介入放射学， 2005， 硕士

【摘要】 目的:运用血管介入插管技术,经犬肾动脉快速注射含人肝细胞生长因子(HGF)重组基因的质粒(pCMV-HGF)。检测不同时间犬肾脏组织中外源性人HGF蛋白的水平,探讨了解外源性HGF基因在犬肾脏组织内的表达情况。 方法:健康成年犬6只,在麻醉和无菌条件下均经肾动脉介入插管。其中4只注射pCMV-HGF质粒,1只注射对照空质粒(pcDNA3),1只注射等渗盐水为无质粒空白对照。应用抗人HGF单克隆和多克隆抗体,建立检测人重组HGF的酶联免疫检测(ELISA)系统。注射后不同时间留取犬静脉血以及部分肾组织,采用ELISA方法,分别检测血清及肾组织中人HGF蛋白的水平。 结果:(1)本实验建立的检测人重组HGF的ELISA敏感性高(15～1000ng/ml),重复性强;(2)接受3种不同处理的犬静脉血中均未检测到人重组HGF蛋白的存在;(3)注射pCMV-HGF质粒的犬肾组织中能够检测到人重组HGF蛋白的存在。72小时肾组织中HGF的水平如表2所示,范围在27～667(ng/mg蛋白)之间。注射空质粒和等渗盐水的犬(即2号和5号犬)肾组织则无表达。 结论:运用血管介入插管技术,经犬肾动脉快速注射含人肝细胞生长因子(HGF)重组基因的质粒(pCMV-HGF)可以在肾脏内产生基因高转染表达。更多还原

【Abstract】 Objective: To evaluate the efficiency of gene transfer and expression by intrarterial injecting naked plasmid encoding human hepatocyte growth factor (HGF).Methods: Six canines were divided into study and control group. Naked plasmid encoding HGF was administrated in the artery of the left kidney in the four canines (study group), while control empty vector was injected in an identical manner in the other two (control group). Groups of canines were sacrificed at day 3 after injection procedure, respectively, and the left kidneys were removed for ELISA.Results: Human HGF protein was detected in the tissue of the left kidneys of the study group, but no detectable human HGF protein was found in left kidneys of control group.Conclusion: Intrarterial administration of naked plasmid encoding human HGF can be an efficient method to express human HGF protein locally.更多还原