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CBP基因沉默对大鼠颈动脉球囊损伤后内膜增生的影响

ISSN号：1007-3949
期刊名称：中国动脉硬化杂志
时间：2013
页码：6-10
分类：R543.3[医药卫生—心血管疾病;医药卫生—临床医学;医药卫生—内科学]
作者机构：[1]三峡大学第一临床医学院心内科三峡大学心血管病研究所,湖北省宜昌市443003, [2]武汉大学人民医院心内科,湖北省武汉市430060
相关基金：国家自然科学基金资助项目（81200088和30770849）;湖北省自然科学基金资助项目（2011CDB179）
相关项目：基因转染RP105抑制TLR4对心肌缺血再灌注损伤的保护及机制研究

关键词： CREB结合蛋白, 基因沉默, 内膜增生, 核因子κB, 乙酰化, CREB Binding Protein, Gene Silencing, Neointimal Hyperplasia, Nuclear Factor KappaB p65, Acetylation

中文摘要：

目的观察小干扰RNA（siRNA）重组慢病毒介导的CREB结合蛋白（CBP）基因沉默对大鼠颈动脉球囊损伤后新生内膜增生的影响，并探讨其作用机制。方法雄性SD大鼠48只，随机分为4组：假手术组、PBS对照组、慢病毒介导CBP基因siRNA转染组（CBP—siRNA—Lenti组）及慢病毒介导非CBP同源序列siRNA转染组（NC-siRNA．Lenti组）。建立大鼠颈动脉球囊损伤模型，术后28天处死动物。分别用实时定量PCR、WesternBlot检测大鼠颈动脉CBP和乙酰化核因子KBp65（NF—κBp65）的表达水平；病理组织学观察血管内膜增生情况；免疫组织化学染色对损伤血管壁增殖细胞核抗原（PCNA）的表达进行评估。结果术后28天，与PBS对照组和NC－siRNA—Lenti组比较，CBP．siRNA—Lenti组CBPmRNA和蛋白的表达显著下调（P均〈0．05），CBP沉默能明显抑制新生内膜面积（0．108±0．008mm2比0．238±0．022mm2、0．252±0．016mm2，P〈0．05）、内膜与中膜面积比（0．706±0．062比1．483±0．136、1．497±0．137，P〈0．05）的增加，及下调血管壁乙酰化NF—κBp65和PCNA的表达水平（P均〈0．05）。结论慢病毒介导的CBP基因沉默能有效地抑制颈动脉球囊损伤后新生内膜的形成，其机制可能与抑制NF—κBp65的过度乙酰化有关。

英文摘要：

Aim To investigate the effects of siRNA recombinant lentivirus targeting CREB binding protein （CBP） gene on neointimal hyperplasia in rat carotid arteries after balloon injury and its possible mechanism. Methods Forty-eight male SD rats were randomly divided into four groups： Sham group, PBS group, CBP-siRNA-Lentivirus group and NC-siRNA-Lentivirus group to establish the model of balloon-injury in left common carotid artery, and the rats were sacrificed 28 days after injury and in vivo gene transfer. The expression of CBP and acetylated nuclear factor kappaB p65 （NF-KB p65） were determined by real-time PCR and Western Blot. Proliferating cell nuclear antigen （PCNA） protein expression was detected by immunohistochemistry. Meanwhile, morphometric analysis was used to measure neointimal hyperplasia. Results 28 days after operations, lentivirus siRNA targeting CBP markedly decreased CBP mRNA and protein expression compared with PBS and NC-siRNA-Lentivirus groups （ P 〈 0. 05 ）. CBP gene silencing significantly reduced the neointimal area （0. 108±0. 008 mm2 vs 0. 238 ±0. 022 mm2 and 0. 252 ±0. 016 mm2, P 〈0. 05） and intima / media ratio （0. 706 ± 0. 062 vs 1. 483 ± 0. 136 and 1. 497 ± 0. 137, P 〈 0. 05 ）. Furthermore, compared with PBS and NC-siRNA-Lentivirus groups, PCNA and acetylation of NF-κB p65 expression were both obviously downregulated in CBP-siRNA-Lentivirus group （P 〈 0. 05 ）. Conclusions Lentivirus-mediated CBP gene silencing could efficiently suppress neointimal formation in balloon injured rat carotid artery, and the mechanism was involved with downregulation of NF-κB p65 acetylation.

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