Han Yaling, Guo Liang, Yan Chenghui, Guo Peng, Deng Jie, Mai Xiaoyan, Kang Jian, Li Shaohua
Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, China.
J Vasc Surg. 2008 Jul;48(1):201-9. doi: 10.1016/j.jvs.2008.01.061. Epub 2008 May 9.
This study examined the effect on neointimal hyperplasia of adenovirus-mediated delivery of cellular repressor of E1A-stimulated genes (CREG) to the artery after balloon injury.
Sixty rabbits were randomized into three groups and underwent balloon injury in the left common carotid arteries. The injured arterial segment was isolated by two inflated balloon catheters. Saline or recombinant adenovirus expressing CREG or green fluorescent protein was injected into the lumen of the isolated arterial segments and incubated for 30 minutes. The rabbits were euthanized for immunohistochemistry, Western blotting, and morphometric analysis at 3, 7, 14, and 28 days after balloon injury and in vivo gene transfer (5 rabbits for each time point). Common carotid artery angiography was performed before euthanasia.
Immunohistochemistry and Western blot analysis demonstrated that CREG expression was significantly down-regulated in the acute phase of vascular injury and was gradually restored in the resolution phase. The changes of CREG expression were in parallel with those of the smooth muscle cell (SMC) differentiation markers SM alpha-actin and SM myosin heavy chain in the injured arteries. Adenovirus-mediated CREG transfer markedly increased CREG expression in the injured artery. Consequently, morphometric analysis revealed an approximate 50% reduction in the neointima and the intima/media ratio in CREG-transferred arteries compared with the saline and green fluorescent protein controls. Assay with 5-bromo-2-deoxyuridine showed that CREG transfer significantly inhibited SMC proliferation. In contrast, endothelialization of the injured artery was not affected by CREG transduction as assessed by CD31 immunostaining.
These data suggest that forced expression of CREG in the artery wall after acute vascular injury inhibits SMC proliferation, induces cellular differentiation, and attenuates neointimal hyperplasia. CREG delivery may have therapeutic potential for the prevention of restenosis after vascular angioplasty.
本研究检测腺病毒介导的E1A刺激基因的细胞抑制因子（CREG）导入球囊损伤后的动脉对新生内膜增生的影响。
60只兔子随机分为三组，对其左颈总动脉进行球囊损伤。用两个充气的球囊导管隔离损伤的动脉段。将生理盐水或表达CREG或绿色荧光蛋白的重组腺病毒注入隔离动脉段的管腔，并孵育30分钟。在球囊损伤和体内基因转移后3、7、14和28天对兔子实施安乐死，用于免疫组织化学、蛋白质印迹和形态计量分析（每个时间点5只兔子）。在安乐死之前进行颈总动脉血管造影。
免疫组织化学和蛋白质印迹分析表明，CREG表达在血管损伤急性期显著下调，在消退期逐渐恢复。CREG表达的变化与损伤动脉中平滑肌细胞（SMC）分化标志物平滑肌α-肌动蛋白和平滑肌肌球蛋白重链的变化平行。腺病毒介导的CREG转移显著增加了损伤动脉中CREG的表达。因此，形态计量分析显示，与生理盐水和绿色荧光蛋白对照组相比，转导CREG的动脉新生内膜和内膜/中膜比值降低了约50%。5-溴-2-脱氧尿苷检测显示，CREG转移显著抑制SMC增殖。相反，通过CD31免疫染色评估，损伤动脉的内皮化不受CREG转导的影响。
这些数据表明，急性血管损伤后动脉壁中CREG的强制表达抑制SMC增殖，诱导细胞分化，并减轻新生内膜增生。CREG递送可能对预防血管成形术后再狭窄具有治疗潜力。
