脂质体介导血管内皮生长因子基因促进大鼠皮瓣成活的实验研究

目的:探讨脂质体介导血管内皮生长因子基因转染大鼠皮瓣,对皮瓣成活和断蒂时间的影响。方法:将30只SD大鼠作为实验模型,随机分为三组,每组10只,制作背部随意皮瓣,分别注入脂质体包裹的PcDNAVEGF165(目的基因组)、PcDNA(空白质粒组)、NS(生理盐水组)。术后3d断蒂,断蒂后7d处死大鼠观察:(1)断蒂后各组皮瓣成活率比较。(2)取皮瓣组织标本常规HE染色,检测平均微血管数目及内径。(3)取皮瓣组织标本行VEGF免疫组化染色检测VEGF表达情况。(4)取皮瓣组织标本在透射电镜下观察超微结构。结果:目的基因组皮瓣平均成活率明显高于空白质粒组和生理盐水组两对照组,平均血管数目明显多于两对照组,目的基因组皮瓣远端血管数目多于近端,免疫组化染色深度明显高于两对照组,均具有统计学意义(P<0.05)。超微结构可见VEGF基因转染组有新生血管形成,内皮细胞内可见较多粗面内质网、线粒体等结构,组织内成纤维细胞增多,细胞合成代谢旺盛。结论:局部注射VEGF基因后可以增加缺血皮瓣血管新生,提高皮瓣成活率,缩短带蒂时间,是一种相对简单、高效的治疗方法。

The Role of Vascular Endothelial Growth Factor Gene in the Experimental Study of Pedicle Flap in Rats

Objective:To explore the effects of liposome-mediated vascular endothelial growth factor(VEGF) transfection on survival and early pedicle division in rats flap.Methods:Divided 30 rats to three teams,after the model of flap was designed in the rats.Purpose gene team inject PcDNAVEGF165,Bland plasmid team inject PcDNA,Physiological saline team inject NS.After flap formed and drug used,We devise the pedical at 3 days.7 days after pedicle division,the rats were killed.Then to measure the flap survival rate.The microvessels was studay by histologic examination.The expression of VEGF was assessed by immunohistochemical staining.The flaps were also examined under the electron ultrastructure microscopy.Results:(1)The flap survival rates were significant different between experimental group and the control group(P<0.05).(2)Micro vascular density and diameter compared PcDNAVEGF165 team with the control teams,there is significant different(P<0.05).(3) Inwrunity histochemistry staining documented were deaper in the PcDNAVEGF165 team than control group,there is significant different between control group.(4)Ultrastructures detection there were new vascular in VEGF transfewed team,endoehelial cell has many rough endoplasmic reticulum,mitoch-ondria etal,lots of fibroblast cells which forms many rough endoplasmic rericnlum has many ribophoring on it,anabolism is bloom.Conclusion:Local injection of VEGF gene can increase angiogenesis in ischemic skin flap to improve flap survival rate,shorten the time pedicle is a relatively simple and efficient method of treatment.