犬经皮插管肾动脉注射人HGF基因的实验研究

Intrarterial administration of naked plasmid encoding human hpatocyte growth factor in canines

目的:运用介入插管技术,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鄄HGF)。检测不同时间犬肾脏组织中外源性人HGF蛋白的水平,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左肾动脉介入插管。其中4只注射pCMV鄄HGF质粒,1只注射空质粒(pcDNA3)对照,1只为注射等渗盐水空白对照。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该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鄄HGF质粒的犬肾组织中能够检测到人重组HGF蛋白的存在。72h肾组织中HGF的水平范围在27～667ng/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:运用介入插管技术,经犬肾动脉快速注射pCMV鄄HGF可以在肾脏内产生基因高转染表达。

Objective: To evaluate the efficiency of gene transfer and expressio n by intrarterial injecting naked plasmid encoding human hepatocyte growth facto r (HGF). Methods: Six canines were divided into the study and the control group. Naked plasmid encoding HGF was administrated in the artery of the left kidney i n the four canines (study group), while control empty vector or saline was injec ted in an identical manner in the other two (control group). Groups of canines w ere sacrificed 3 days after injection procedure, respectively, and the left kidn eys were removed for ELISA. Results: Human HGF protein was detected in the tissu e of the left kidneys of the study group, but no detectable human HGF protein wa s found in left kidneys of the control group. Conclusion: Intrarterial administr ation of naked plasmid encoding human HGF can be an efficient method to express human HGF protein locally.