摘要
目的探讨以脂质体和腺病毒为载体,早期生长反应基因-1 DNA酶(Egr-1 DNA enzyme,EDRz)局部外用对自体移植静脉血管平滑肌(VSMC)增殖和内膜增生的抑制作用。方法大鼠建立自体静脉移植模型后随机分为脂质体组、腺病毒组和对照组,两实验组分别用脂质体-EDRz和腺病毒-EDRz涂抹移植静脉行在体转染。于术后1,2,6,24 h及3,7,14,28 d取材,荧光显微镜检测移植静脉的转染情况;采用原位杂交方法检测Egr-1 mRNA的表达;用免疫组织化学方法检测Egr-1蛋白表达,同时进行组织形态学观察。结果术后1 h,EDRz主要位于移植静脉的外膜、中膜和部分内皮细胞。脂质体组荧光灰度值为70.3±13.5,腺病毒组荧光灰度值为60.5±11.2。术后2～24 h,EDRz主要位于移植静脉的中膜;移植7 d,EDRz主要位于移植静脉的内膜。移植早期Egr-1蛋白表达主要位于中膜的血管平滑肌细胞(VSMC)和部分单核细胞、内皮细胞;移植后期未检测到Egr-1 DNA酶的表达。移植后期在中膜和新生内膜的VSMC均未检测到Egr-1蛋白的表达。移植2 h阳性表达率,脂质体组为(15.3±4.2)%,腺病毒组为(20.7±2.6)%。脂质体组和腺病毒组与对照组比较,前两者VSMC的增殖程度和内膜厚度明显减轻。结论 EDRz通过减少Egr-1在自体移植静脉中的表达,可有效地抑制自体移植静脉中VSMC的增殖和内膜增生;脂质体转染优于腺病毒转染。
Objective To investigate the inhibitory effects of the liposome-and adenovirus-mediated early growth response gene-1 DNA enzyme（EDRz） through regional external application on the vascular smooth muscle cells（VSMCs） proliferation and intimal hyperplasia of the autogenous vein graft.Methods The autogenous vein graft model was established in rats,which were then randomly divided into liposome group,adenovirus group and control group.In vivo transfection was conducted in the two experimental groups by regional application of liposome-EDRz and adenovirus-EDRz on the vein graft,respectively.The vein graft samples were harvested at 1,2,6 and 24 hours,and also 3,7,14,28 and 42 days after surgery,respectively.The transfection status of vein graft was detected with fluorescent microscope,Egr-1 mRNA and protein expression was measured by in situ hybridization and immunohistochemical staining,respectivcely.Meanwhile,the histomorphology of the vein graft was also observed.Results At one hour after surgery,EDRz was mainly located in media,adventitia and part of endothelial cells of the vein graft.The fluorescent intensity of liposome group and adenovirus group was 70.3±13.5 and 60.5±11.2,respectively.In 2 to 24 h after surgery,EDRz was located in the media of the vein graft,and it was mainly located in the intima of the vein graft at 7 h after surgery.No EDRz was detected in the vein grafts at the later postoperative period.In the early postoperative period,Egr-1 protein mainly expressed in the medial VSMCs,and in some monocytes and endothelial cells,but it could not be detected in either the media or neointimal VSMCs of the vein graft at the later postoperative period.At 2 h after surgery,the positive expression rate of Egr-1 protein of liposome group and adenovirus group was（15.3±4.2）% and（20.7±2.6）%,respectively.The degree of proliferation of VSMC and intima thickness of liposome group and adenovirus group were obviously decreased compared with control group.Conclusions EDRz has inhibitory effect on VSMCs proliferation and intimal hyperplasia of the autogenous vein graft through its Egr-1 expression.Liposome transfection is more efficient than adenovirus transfection.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2011年第12期1336-1341,共6页
China Journal of General Surgery
基金
国家自然科学基金资助项目(30801123
30872527)
