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腺病毒介导的AT2R基因转染对大鼠颈动脉新生内膜增生的抑制作用 被引量：1

Inhibition of neointimal hyperplasia after balloon angioplasty of carotid artery with transfection of adenovirus-mediated AT2R gene in rat
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摘要 目的构建AngⅡ2型受体(AT2R)重组腺病毒载体,并研究AT2R基因转染对大鼠颈动脉新生内膜增生的影响。方法用细菌内同源重组方法构建带有绿色荧光蛋白(GFP)报告基因的AT2R重组腺病毒载体pAdCMV/AT2R。大鼠颈动脉球囊损伤后,将pAdCMV/AT2R或空病毒pAdGFP局部导入,分别用RTPCR、免疫荧光染色、激光共聚焦技术检测目的基因AT2RmRNA及蛋白在血管中的表达,利用图像分析仪进行组织形态学分析。结果重组腺病毒载体pAdCMV/AT2R经PCR鉴定正确,其滴度为1.5×1012pfu/ml。pAdCMV/AT2R局部转染后14天,大鼠颈动脉新生内膜、外膜、中层有绿色荧光,阳性面积约40%;转染后7、14、21天大鼠颈动脉AT2RmRNA及蛋白表达显著高于pAdGFP转染组,21天时其内膜与中层的面积比显著低于pAdGFP组(0.78±0.06vs1.36±0.21,P<0.01)。结论pAdCMV/AT2R在体转染能上调血管中AT2R的表达,显著抑制大鼠颈动脉球囊损伤后新生内膜增生。 Objective the construct the recombinant adenovirus of Angiotensin Ⅱ type 2 receptor （AT2R）. and study the effect of AT2R on prevention of neointimal hyperplasia in rat carotid arteries after balloon angioplasty. Methods The recombinant adenovirus of AT2R with green fluorescent protein was constructed by using the method of homogenous recombination in bacteria. AT2R gene was transduced into rat carotid arteries with pAdCMV/AT2R after the reproduction of rat carotid balloon injury restenosis model. The expression and the transfection efficiency of AT2R gene were evaluated by RTPCR, immunofluorescence staining, and confocal microscope. The intimal/medial area ratio was measured by digital analysis system. Results The titre of purified recombinant adenovirus was 1.5 × 10^12 pfu ml, and it was proved by PCR. pAdCMV/AT2R transfection efficiency in carotid artery was about 40% at day14, and localized to the neointima, as well as to the media and the adventitia, pAdCMV/AT 2R delivered into injured rat carotid arteries significantly up-regulated AT2RmRNA and protein expression in neointima from day 7 to 21 after injury. Compared with pAd GFP transfection group, pAdCMV AT2R transfection reduced I/M ratio significantly on day 21（0.78±0.06 vs. 1.36±0.21 respectively, P〈0.01）. Conclusion Adenovirus-mediated gene transfer of AT2R significantly up-regulate AT 2RmRNA and protein expression in neointima and effectively inhibit neointimal hyperplasia in the rat carotid arteries after balloon angioptasty. It may potentially be developed as a new approach to prevent restenosis after angioptasty.
作者 唐兵 何国祥 李德 刘建平 景涛
机构地区 第三军医大学西南医院心内科
出处 《解放军医学杂志》 CAS CSCD 北大核心 2005年第12期1068-1071,共4页 Medical Journal of Chinese People's Liberation Army
关键词 受体 血管紧张素 2型 转染 血管内膜 receptor, angiotensin, type 2 transfection tunica intima
分类号 R543.4 [医药卫生—心血管疾病]
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参考文献9

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二级参考文献7

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共引文献1

同被引文献9

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解放军医学杂志
2005年 第12期

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