大鼠颈动脉球囊损伤局部A20基因转染后内皮细胞再生及血管超微结构的影响  

Effects of zinc finger protein A20 on reendothelialization and vascular ultrastructure of rats with carotid artery balloon injury

作　　者：高静[1] 李蕾[1] 徐丹[1] 陈良[1] 曲鹏[1] 

机构地区：[1]大连医科大学附属第二医院心内科,辽宁省大连市116027

出　　处：《中国组织工程研究与临床康复》2008年第15期2838-2842,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30670836)~~

摘　　要：目的：锌脂蛋白A20是核因子κB信号系统活化的产物。研究证实核因子κB的激活是球囊损伤动脉后血管狭窄发生的重要机制之一。实验拟观察锌指蛋白A20对大鼠颈总动脉球囊损伤局部血管内皮再生和血管超微结构的影响。方法：实验于2006-03／2007-06在大连医科大学附属第二医院中心实验室完成。①实验材料：健康雄性SD大鼠42只，体质量350-400g。②分组及实验过程：采用随机数字表法将大鼠分为3组，每组14只。制备A20质粒DNA，建立大鼠颈总动脉球囊损伤模型（假手术组只进行颈动脉结扎，不进行球囊损伤术）；对照组：球囊损伤术后局部灌注Lipofectamine 2000 Reagent＋TE缓冲液；治疗组：球囊损伤术后局部灌注Lipofectamine 2000 Reagent＋TE缓冲液＋pCAGGS-GFP／A20重组质粒。A20质粒DNA在球囊损伤大鼠颈动脉的局部转染。③实验评估：荧光显微镜观察A20在损伤动脉壁的转染效率；伊文思蓝染色法评估损伤后颈动脉内皮再生情况；透射电镜观察血管超微结构改变。实验过程对动物的处理符合动物论理学标准。结果：①球囊损伤术和A20转染治疗后2周，治疗组再内皮化百分比大于对照组（P〈0．05）。②假手术组大鼠颈动脉平滑肌细胞电镜下呈典型的“收缩表型”；对照组球囊损伤后3d可见合成型细胞及过渡型细胞，7d血管平滑肌细胞表型发生明显改变，呈典型的“合成表型”，治疗组术后7d可见接近收缩型的血管平滑肌细胞。结论：局部转染A20基因可促进损伤血管内皮再生，抑制损伤处血管平滑肌细胞表型转化。AIM： Zinc finger protein A20 is the activated product of nuclear factor- κB signaling. Activation of nuclear factor- κB is one of important mechanisms of angiostegnosis after balloon injury. This study aimed to investigate the effects of in vivo local transfection of zinc finger protein A20 gene on reendothelialization and vascular ultrastructure of rats with carotid artery balloon injury. METHODS： Experiments were performed at the Central Laboratory of Second Affiliated Hospital of Dalian Medical University from March 2006 to June 2007.（1）Forty-two healthy male Sprague-Dawley （SD） rats of 350-400 g were selected. （2）The rats were randomly divided into three groups equally. A20 plasmid DNA was prepared. Rat models of common carotid artery balloon injury were established. Rats in the sham operation group only received ligation of carotid. Rats in the control group were administered with Lipofectamine 2000 Reagent＋TE buffer solution in local region after sac injury. Rats in the treatment group were treated with Lipofectamine 2000 Reagent ＋TE buffer solution＋pCAGGS-GFP/A20 recombinant plasmid after sac injury. A20 plasmid DNA was locally transferred into rats with carotid balloon injury. （3）The transfection efficiency of A20 plasmid in injured arterial wall was evaluated by fluorescence microscope. Endothelial regeneration of corotid artery was evaluated by Evans blue dye staining. Ultrastructural change in injured artery was observed with a transmission electron microscope. The disposal in animals was accorded with the animal ethical standards. RESULTS： （1）Local transfection of A20 gene resulted in an increase in reendothelialization, as measured as percent area reendothelialized at day 14 in the treatment group compared to the control group （P 〈 0.05）. （2）Contractile phenotype of vascular smooth muscle cells （VSMCs） was shown in the sham operation group and synthetic phenotype and transition cells were shown after balloon injury in the control group at day 3. VSMCs s

关 键 词：锌指蛋白A20 内皮 超微结构 基因转染 大鼠 

分 类 号：R329.412[医药卫生—人体解剖和组织胚胎学]