Local matrix metalloproteinase 2 gene knockdown in balloon-injured rabbit carotid arteries using nonviral-small interfering RNA transfection
Local matrix metalloproteinase 2 gene knockdown in balloon-injured rabbit carotid arteries using nonviral-small interfering RNA transfection
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DOI：
10.1002/jgm.1275
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发表时间：
2009-01-01
影响因子：
3.5
通讯作者：
Feldman, Laurent J.
中科院分区：
文献类型：
--
作者：
Hlawaty, Hanna;Juan, Aurhe San;Feldman, Laurent J.
背景小干扰RNA（siRNA）是治疗心血管疾病的一种很有前途的方法。基质金属蛋白酶（MMP）2在动脉壁中的过度表达与经皮冠状动脉介入治疗后的再狭窄以及自发性动脉粥样硬化斑块破裂有关。我们假设体内局部递送靶向MMP 2的siRNA，方法球囊损伤后2周，5 μ mol/l Tamra标记的MMP 2-siRNA，将scramble siRNA或生理盐水局部注射到颈动脉中，孵育1 h。siRNA在新生内膜细胞的表层。通过实时逆转录聚合酶链反应测定的MMP 2 mRNA水平在MMP 2-siRNA转染的动脉中比scramble siRNA转染的动脉中降低了79 +/- 25%（p < 0.05）。通过明胶酶谱法在MMP 2-siRNA与scramble siRNA转染的动脉的条件培养基上进行测量，MMP 2活性在24、48和72小时分别降低了53 +/-29%、50 +/- 24%和46 +/- 14%（所有p < 0.005）。MMP 9，pro-MMP 9和TIMP-2 levels.Conclusions在本研究中获得的结果表明，显着的基因表达的抑制，可以实现与局部交付的siRNA在体内动脉壁。版权所有（C）2008约翰威利父子有限公司
Background Small interfering RNA (siRNA) delivery is a promising approach for the treatment of cardiovascular diseases. Matrix metalloproteinase (MMP) 2 over-expression in the arterial wall has been implicated in restenosis after percutaneous coronary intervention, as well as in spontaneous atherosclerotic plaque rupture. We hypothesized that in vivo local delivery of siRNA targeted at MMP2 (MMP2-siRNA) in the balloon-injured carotid artery of hypercholesterolemic rabbits may lead to inhibition of MMP2 expression.Methods Two weeks after balloon injury, 5 mu mol/l of Tamra-tagged MMP2-siRNA, scramble siRNA or saline was locally injected in the carotid artery and incubated for 1 h.Results Fluorescent microscopy studies showed the circumferential uptake of siRNA in the superficial layers of neointimal cells. MMP2 mRNA levels, measured by the real-time reverse transcriptase-polymerase chain reaction, was decreased by 79 +/- 25% in MMP2-siRNA- versus scramble siRNA-transfected arteries (p < 0.05). MMP2 activity, measured by gelatin zymography performed on the conditioned media of MMP2-siRNA versus scramble siRNA transfected arteries, decreased by 53 +/- 29%, 50 +/- 24% and 46 +/- 14% at 24, 48 and 72 h, respectively (p < 0.005 for all). No effect was observed on MMP9, pro-MMP9 and TIMP-2 levels.Conclusions The results obtained in the present study suggest that significant inhibition of gene expression can be achieved with local delivery of siRNA in the arterial wall in vivo. Copyright (C) 2008 John Wiley & Sons, Ltd.