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Egr-1的诱骗性寡脱氧核苷酸抑制大鼠颈总动脉损伤后MMP-2的表达
引用本文:孙晓花,刘闺男,张晶.Egr-1的诱骗性寡脱氧核苷酸抑制大鼠颈总动脉损伤后MMP-2的表达[J].中国组织化学与细胞化学杂志,2013(5):366-371.
作者姓名:孙晓花  刘闺男  张晶
作者单位:中国医科大学附属第一医院心血管内科,沈阳110001
基金项目:国家自然科学基金项目资助(30871074)
摘    要:目的观察局部转染早期生长反应因子-1(early growth response factor-1,Egr-1)的特异诱骗寡脱氧核苷酸(decoy oligodeoxynucleotides,decoy ODNs)对球囊损伤颈总动脉后基质金属蛋白酶-2(MMP-2)蛋白表达的影响及内膜增生的情况,初步探讨Egr-1,decoy ODNs抑制球囊损伤后内膜增生的机制。方法 96只健康雄性Wistar大鼠,随机分为4组,分别为假手术组、对照组、杂码组和诱骗组,每组24只。除假手术组外均应用2F球囊导管行颈总动脉球囊损伤术,术中采用转染试剂FuGENE6介导的Egr-1decoy ODNs转染至损伤后大鼠血管中,与假手术组、对照组、杂码组相比较。术后3、7、14、21d每组处死6只动物。应用HE染色和免疫组织化学方法观察大鼠颈总动脉球囊损伤后内膜增生情况和MMP-2蛋白的表达及转染Egr-1decoy ODNs后对它们的影响。结果 (1)、内膜损伤后3d内膜增厚不明显,7d内膜开始增厚,14、21d时内膜明显增厚。(2)、在假手术组近腔面中膜可见MMP-2有少量散在阳性表达;在对照组及杂码组动脉损伤后3d,在近腔面中膜,有少量阳性表达,与假手术组相比,阳性表达指数上升。7d时在新生内膜和靠近新生内膜处中膜表达明显,14d后表达逐渐下降。(3)转染decoy ODNs治疗后,在各个时间点内膜增厚程度减轻,MMP-2蛋白表达减少,与对照组比较差异有显著性(P<0.01)。结论血管球囊损伤后,内膜7d开始增生,14d、21d增生更明显,而MMP-2在7d时表达明显,之后逐渐下降,Egr-1decoy ODNs能抑制MMP-2的表达,从而减轻血管损伤后内膜的增生。

关 键 词:球囊损伤  早期生长反应因子-1  诱骗性寡脱氧核苷酸  MMP-2  血管内膜增生

Egr-1 Decoy oligodeoxynucleotides Down-regulate MMP-2 after Balloon Injury of Rat Common Carotid Artery
Sun Xiaohua,Liu Guinan,Zhang Jing.Egr-1 Decoy oligodeoxynucleotides Down-regulate MMP-2 after Balloon Injury of Rat Common Carotid Artery[J].Chinese Journal of Histochemistry and Cytochemistry,2013(5):366-371.
Authors:Sun Xiaohua  Liu Guinan  Zhang Jing
Institution:(Department of Cardiology, The First Affiiliated Hospital of China Medical University ,Shenyang 110000, China)
Abstract:Objective To observe the effects of the local transfection of decoy ODNs (decoy oligodeoxynucleotides) targeting Egr-1 (early growth response factor-1) on matrix metalloproteinase-2 (MMP-2) expression and intimal hyperplasia after artery balloon injury, and to explore its possible mechanism of inhibiting neointimal formation. Methods 96 healthy male Wistar rats were randomly divided into 4 groups: sham operation group, control group, miscellaneous code group and decoy group, with 24 rats each. Except the sham operation group, the other groups underment carotid artery balloon injury. Intraoperative use of transfection reagent FuGENE6 transfected Egr-1 decoy ODNs into the injured blood vessels of the decoygroup. After 3, 7, 14 and 21d, 6 rats of each group were killed. He staining and immunohistochemi- cal method were used to observe the influence of balloon injury of rat carotid artery and transfeetion of Egr- 1 decoy ODNs on intimal hyperplasia and the expression of MMP-2 protein. Results (1)The intimal thick- ness existed at day 7 in control and miscellaneous code group and it was more significant at day 14 and 21. (2)The expressions of MMP-2 were at low levels in medial layers near the lumen in control and miscellous groups at day 3, reached their peaks in neointimal and medial layers at day 7 and began decreasing at day 14 after injury. (3)The level of MMP-2 was reduced and the intimal hyperplasia was inhibited after treatment with Egr-1 decoy ODNs. Compared with sham-operation group and control group, there were significant differ ences (P〈0. 01). Conclusion After vascular balloon injury, intimal hyperplasia begins at 7d, and becomes more obvious at 14d and 21d. MMP-2 expression is significant at 7d and then decreases gradually. Egr-1 decoy ODNs can inhibit MMP-2 expression, thus reducing the intimal hyperplasia after vascular injury.
Keywords:Balloon injury  Egr-1  Decoy ODN  MMP-2  Neointimal hyperplasia
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