肌肉注射含内皮PAS结构域蛋白1(EPAS1)基因的慢病毒改善外周动脉血管病模型大鼠后肢缺血及其机制  

Intramuscular injection of lentivirus-mediated EPAS1 gene improves hind limb ischemia and its mechanism in a rat model of peripheral artery vascular disease

作　　者：王志鸿[1] 顾洪斌[1] 杨帆[1] 解华杰 盛磊[1] 李明飞[1] 

机构地区：[1]解放军306医院血管外科,北京100101

出　　处：《细胞与分子免疫学杂志》2017年第11期1539-1543,共5页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的研究内皮PAS结构域蛋白1(EPAS1)过表达对外周动脉病大鼠模型的保护作用。方法利用单侧髂外动脉结扎法制备大鼠外周动脉血管病模型,将带有EPAS1基因的慢病毒注射到大鼠右侧大收肌。利用步态分析仪检测大鼠步态改变,激光多普勒检测大鼠后肢血流量变化,实时定量PCR检测EPAS1和促血管生长的肝细胞生长因子(HGF)、碱性成纤维细胞生长因子(bFGF)和血管内皮细胞生长因子(VEGF)的mRNA水平,免疫组织化学染色检测α平滑肌肌动蛋白(αSMA)水平。结果成功制备了大鼠后肢缺血模型,与注射对照病毒lenti-EGFP组相比,从手术后7 d开始,lenti-EPAS1注射组大鼠步态障碍得到明显缓解,后肢血流量明显恢复;缺血大鼠大收肌中HGF、bFGF、VEGF的mRNA水平增加,肌组织中αSMA表达明显增多。结论过表达EPAS1能改善大鼠缺血部位血流量,促进相关细胞因子表达和新生血管生成。Objective To investigate the effect of over-expressed endothelial Per-Arnt-Sim domain protein 1（EPAS1） on peripheral arterial disease（ PAD） in a rat model. Methods PAD rat model was established by external iliac artery ligation followed by lentivirus-mediated EPAS1 gene injection into rat right adductor magnus. The models were evaluated by quantitative analysis of gait disturbance. The changes of blood flow in the posterior extremity of the rats were detected using laser Doppler. The expressions of EPAS1,hepatocyte growth factor（ HGF）,basic fibroblast growth factor（ bFGF）,and vascular endothelial growth factor（ VEGF） mRNAs were tested by real-time quantitative PCR. The expression of α-smooth muscle actin（ αSMA） was detected by immunohistochemical staining. Results Compared with lenti-EGFP group,rat hind limb function and circulation got recovered obviously 7 days after lenti-EPAS1 injection. The mRNA expressions of EPAS1,HGF,bFGF,and VEGF were up-regulated in the lenti-EPAS1-treated sites. The expression of αSMA showed an obvious increase in the lenti-EPAS1-treated muscles. Conclusion Over-expressed lenti-EPAS1 can promote angiogenesis via the up-regulation of EPAS1-related angiogenic factors in the muscles of the affected hind limb and reduce gait disturbance.

关 键 词：外周动脉疾病 间歇性跛行 血管生成 步态障碍 内皮PAS1蛋白EPAS1 

分 类 号：R543[医药卫生—心血管疾病]