基因枪在大鼠闭塞性血管病基因治疗中的应用  

Application of gene gun in treatment of occlusive vascular diseases in rats

作　　者：哈小琴[1] 苑宾[1] 李元敏[1] 姚山麟 吴祖泽[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]宁波新芝科器研究所

出　　处：《生物技术通讯》2001年第4期266-269,共4页Letters in Biotechnology

摘　　要：探讨利用基因枪技术转移肝细胞生长因子基因治疗大鼠肢体闭塞性血管病的可行性。构建了携带人肝细胞生长因子 (HGF)基因的重组真核表达载体 (pUDKH)。在制备好大鼠下肢闭塞性血管病模型后 ,通过基因枪或肌肉直接注射法 ,向局部缺血部位肌肉中转移pUDKH ,每只 5 μg(基因枪 )和 12 μg(肌肉注射 )。应用常规组织病理切片 (H .E .染色 )及免疫组织化学方法观察血管形成及基因表达。转移pUDKH后第 10天 ,用基因枪和直接注射法转移的局部肌肉组织内HGF的表达明显高于转移空白质粒 (pUDK)的对照组 ;pUDKH组可见明显的小血管新生 ,而pUDK组至 2 0天时仍未观察到或仅见到极少量的新生血管。基因枪与肌肉注射两组相比血管密度无明显差异。采用基因枪直接转移pUDKH裸露质粒于肢体缺血局部的方法是可行的 ,转移的基因可在局部有效表达 ,达到促进血管形成的预期目的。To explore the feasibility of gene therapy with hepatocyte growth factor (HGF) transferred using gene gun technique in treatment of peripheral vascular disease of rat limb. A model of occlusive vascular disease in rat posterior limb was established and a recombinant vector carrying a human HGF gene (pUDKH) was constructed. pUDKH was transferred into ischemic muscles of rat posterior limb by gene gun (5 μg) direct intramuscular injection (120 μg). The expression of HGF and neovascular formation in the muscle were observed using immunohistochemistry and routine histopathological sections, respectively. On day 10 after gene transfer, the expression level of HGF was evidently higher in local muscles transferred with pUDKH than those with the blank plasmid (pUDK) via both gene gun and intramuscular injection. In muscles transferred with pUDKH, small neovessels were abundant on day 10, whereas in pUDK transferred muscles only a scanty, even none of such blood vessels could be seen on day 20. There was no remarkably difference in blood vessel density of pUDKH transferred muscles between gene gun and intramuscular injection groups. Therefore, direct transfer of naked pUDKH plasmid using gene gun into locally ischemic limb muscles is feasible. The gene transferred in this way can be expressed effectively in eukaryotic cells muscles, so that the expected purpose of promoting neovascular formation would be achieved.

关 键 词：基因枪 肝细胞生长因子 基因治疗 闭塞性血管病 大鼠 

分 类 号：R543.05[医药卫生—心血管疾病] R394.8[医药卫生—内科学]