反义寡核苷酸抑制早期生长反应基因1表达对移植静脉内膜增生的影响  

Effect of transfect early growth response gene-1 antisense oligodeoxynucleotides on the prevention of intimal hyporplasia of autogenous vein graft in rats

作　　者：刘程伟[1] 胡新华[2] 王石[1] 张雪松[1] 王树卿[1] 张强[2] 辛世杰[2] 段志泉[2] 

机构地区：[1]佳木斯大学附属第一医院普外一科,154003 [2]中国医科大学附属第一医院血管外科

出　　处：《中华普通外科杂志》2008年第1期54-57,共4页Chinese Journal of General Surgery

基　　金：国家自然科学基金资助项目（30672048）;黑龙江省高等学校青年学术骨干支持基金资助项目（1152G041）;黑龙江省教育厅科技基金资助项目（11521275）

摘　　要：目的研究反义寡脱氧核苷酸（antisense oligodeoxynucleotides，ASODN）抑制早期生长反应基因1（early growth response gene-1，Egr-1）表达对移植静脉血管平滑肌细胞增殖和内膜增生的影响。方法构建Egr-1 ASODN，建立自体静脉移植模型，Egr-1 ASODN转染移植静脉，术后随机分为1、2、6、24h，3、7、14、28、42d组，以未应用Egr-1 ASODN的大鼠为对照组。荧光显微镜检测Egr-1 ASODN转染移植静脉情况；应用原位杂交和RT—PCR方法检测Egr-1 mRNA的表达；联合应用免疫组织化学方法和Western blot检测Egr-1蛋白表达情况。结果实验组移植1h，Egr-1 ASODN主要位于移植静脉的外膜、中膜和部分内皮细胞（荧光灰度值为67±11），移植2h至24h，Egr-1 ASODN则主要位于移植静脉的中膜，移植7d，Egr-1 ASODN主要位于移植静脉的内膜。移植后期未检测到Egr-1 ASODN的表达。Egr-1 mRNA的表达只呈现一个高峰（基因表达值1．8±0．5）。移植早期Egr-1蛋白表达主要位于中膜的VSMC、部分单核细胞和内皮细胞，而移植后期在中膜和新生内膜的VSMC都未检测到Egr-1蛋白的表达。与对照组相比VSMC的增殖程度和内膜厚度均明显减轻（P〈0．01）。结论Egr-1 ASODN可显著抑制移植静脉的内膜增生，其作用可能是通过抑制Egr-1基因及其蛋白产物表达，从而抑制VSMC增殖、促进其凋亡而实现的。Objective To detect the inhibitory action of early growth response gene-1 antisense oligodeoxynucleotides （Egr-1 ASODN） on vascular smooth muscle cell （VSMC） proliferation and intimal hyperplasia（IH） of autogenous vein graft in rats. Methods Egr-1 ASODN was constructed. Autogenous vein graft model was established in 200 Wistar rats. Egr-1 ASODN was transfected into the graft vein and the vein graft samples were harvested at 1,2,6,24 hours, and 3 .7,14,28,42 days after surgery. Rat without Egr-1 ASODN transfection served as control. Fluorescent microscopy was used to detect the expression of Egr-1 ASODN transfected vein graft. Egr-1 mRNA was measured by RT-PCR and in situ hybridization. Western blotting and immunohistochemistry were used to detect the protein expression of Egr-1. Results At 1 h after grafting in experimental group, Egr-1 ASODN was mainly located in adventitia, media and a fraction of endothelial cells in the vein graft （ fluorescence expression value of 67 ± 11 ）. From 2 h to 24 h Egr-1 ASODN was located in the media of vein graft, and mainly in the intima of vein graft at 7 hr after. There was no Egr-1 ASODN expression in vein grafts at the later phases. Compared with control, the expression of Egr-1 mRNA showed single peak in this study （ gene expression value of 1.8 ± 0.5 ）. We found that Egr-1 protein mainly located in VSMC in the medial. Compared with control the degree of proliferation of VSMC and thickness of intima were relieved （ P 〈 0. 01 ）. Conclusion Transfection of Egr-1 ASODN could inhibit the intimal hyperplasia after vein graft, possibly by a mechanism in which the expression of Egr-1 and proliferation of VSMC were inhibited and cell apoptosis accelerated.

关 键 词：静脉 寡脱氧核糖核苷酸类 反义 早期生长反应基因1 

分 类 号：R686[医药卫生—骨科学]