Elastic fibers reconstructed using adenovirus-mediated expression of tropoelastin and tested in the elastase model of abdominal aortic aneurysm in rats
Elastic fibers reconstructed using adenovirus-mediated expression of tropoelastin and tested in the elastase model of abdominal aortic aneurysm in rats
复制标题
使用腺病毒介导的原弹性蛋白表达重建弹性纤维，并在大鼠腹主动脉瘤弹性蛋白酶模型中进行测试
DOI：
10.1016/j.jvs.2008.04.016
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发表时间：
2008-10-01
影响因子：
4.3
通讯作者：
Ye, Cai Sheng
中科院分区：
文献类型：
--
作者：
Xiong, Jiang;Wang, Shen Ming;Ye, Cai Sheng
目的：将带绿色荧光蛋白（GFP）标签的重组tropoelastin （TE）基因的腺病毒载体转移至主动脉血管平滑肌细胞（VSMCs），研究重组弹性蛋白的体外表达和实验性腹主动脉瘤（AAA）体内弹性纤维的重建。用猪胰腺弹性蛋白酶灌注大鼠动脉诱导动脉瘤形成，形成后将腺病毒载体直接灌注到动脉瘤腔内。用荧光显微镜观察转染AdTE-GFP的VSMCs体外和体内的变化。实时逆转录聚合酶链式反应（RT-PCR）和Western blot法分别比较AdTE-GFP转染VSMCs体外（1、3、5 d）和体内（2、4周）的TE mRNA水平和重组弹性蛋白表达水平。在三个时间点（灌注后立即、2周和4周）测量动脉瘤的主动脉直径（AD），并将转染AdTE-GFP组、空腺病毒（AdNull）组和磷酸缓冲盐水（PBS）灌注组进行比较。通过组织学分析评估体内新弹性纤维的形成。结果。AdTE-GFP转染逆转了AAA的形成（平均标准误差），对照组显示AD增加（PBS灌注组增加48.50 +/- 16.55%，AdNull处理组增加39.84 +/- 15.59%），而AdTE-GFP转染组显示AD减少(23.04 +/- 14.49%，P
Objective: An adenoviral vector carrying a recombinant tropoelastin (TE) gene with a Green Fluorescent Protein (GFP) tag adenoviral tropoelastin green fluorescent protein (AdTE-GFP) was transferred to aortic vascular smooth muscle cells (VSMCs) for studying the expression of recombinant elastin in vitro and the reconstruction of elastic fibers in vivo in experimental abdominal aortic aneurysm (AAA).Methods. The AAAs were induced in rats by perfusing the arteries with porcine pancreatic elastase, and after AAA formation, adenoviral vectors were perfused directly into the aneurysmal lumen. VSMCs transfected with AdTE-GFP in vitro and in vivo were detected by fluorescence microscopy. The TE mRNA levels and the level of recombinant elastin expression of AdTE-GFP transfected VSMCs in vitro (at 1, 3, and 5 days) and in vivo (at 2 and 4 weeks) were compared by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. The aortic diameters (AD) of the aneurysms were measured at three time points (immediately, 2 weeks, and 4 weeks post-perfusion) and comparisons were made among the AdTE-GFP transfected group, an empty adenovirus (AdNull) transfected group, and a phosphate buffered saline (PBS) perfused group. Formation of new elastic fibers in vivo was assessed by histologic analysis. Results. AdTE-GFP transfection reversed AAA formation (mean standard error), with the controls showing increased ADs (a 48.50 +/- 16.55% increase for the PBS perfusion group and a 39.84 +/- 15.59% increase for the AdNull treated group) and the AdTE-GFP transfected group showing a decreased AD (23.04 +/- 14.49%, P