联合促血管生成素-1及血管内皮细胞生长因子基因治疗对大鼠新生血管通透性的影响  

Effects of combined angiopoietin-1 and vascular endothelial growth factor gene transfer on neovascular permeability of rat hindlimb ischemia

作　　者：祖凌云[3] 蒋捷[3] 姜宁 陈莉[4] 郭艳红[2] 李黔[4] 高炜[2] 

机构地区：[1]不详 [2]北京大学第三医院心内科 [3]北京大学第一医院心内科100034 [4]北京大学医学部心血管研究所

出　　处：《中国介入心脏病学杂志》2004年第2期107-110,共4页Chinese Journal of Interventional Cardiology

基　　金：国家自然科学基金资助 (3 0 2 40 0 40 );教育部教育振兴行动计划特殊专项("九八五"工程 )资助

摘　　要：目的　探讨联合转染促血管生成素 1(angoipoietin 1,Ang 1)及血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)基因对大鼠下肢新生血管通透性的影响 ,并观察不同基因剂量配比的差异。方法　制造大鼠下肢血管闭塞病理模型 ,按不同剂量配比肌肉内电转pcD2 Ang 1和 或pcD2 VEGF基因。应用逆转录聚合酶链反应、免疫组化染色检测外源基因在骨骼肌中的表达 ,通过伊文思蓝灌注观察其对新生血管通透性的影响。结果　逆转录聚合酶链反应及免疫组化染色均能检测到外源基因的表达。随着转入pcD2 VEGF剂量的增加 ,血管通透性逐渐增加 ;而随着转入pcD2 Ang 1剂量的增加 ,血管通透性逐渐下降。其中pcD2 VEGF 10 0 μg +pcD2 Ang 12 0 0 μg组通透性与正常对照最为接近 [( 8 5 7± 0 74 )ng mlvs ( 8 4 2± 0 82 )ng ml,P >0 0 5 ]。结论　联合转pcD2 Ang 1基因可逆转pcD2 VEGF基因增加血管通透性的副作用 ,其中pcD2 VEGF与pcD2 Ang 1比例为 1 2时血管通透性最接近生理状态。与单基因治疗相比 ,适当配比的联合转基因可能成为治疗闭塞性血管病更安全、更有效的新方法。Objective To investigate the effect of combined gene transfer of Angiopoietin 1 (Ang 1) gene and Vascular Endothelial Growth Factor (VEGF) gene, with different proportions, on neovascular permeability in a rat model of hindlimb ischemia Methods Naked plasmid DNA encoding VEGF (pcD 2/VEGF) and Ang 1 (pcD 2/Ang 1) were delivered either alone or in combination into the rat ischemic skeletal muscle by intramuscular electroporation Different proportions of the two genes in the combination were investigated RT PCR and immunohistochemistry were used to detect gene expression The permeability of the neovasculature was evaluated by the skeletal muscle Evans blue uptake assay Results High levels of VEGF and Ang 1 mRNA and protein expression were detected Quantification of the leakage levels of Evans blue dye demonstrated that the increase of the amount of pcD 2/VEGF from 100 μg, 200 μg to 400 μg resulted in a gradual increase in the vascular permeability, whereas the vasculas permeability was decreased with the increase of the amount of pcD 2/Ang 1 Evans blue uptake in group of pcD 2/VEGF 100 μg + pcD 2/Ang 1?200 μg was the closest to that of the normal group (8 57±0 74 ng/ml vs 8 42±0 82 ng/ml, P >0 05) Conclusion Intramuscular electroporation of pcD 2/Ang 1 can ameliorate the vessel leakage caused by pcD 2/VEGF and thus may represent a feasible and safe therapeutic strategy for patients with tissue ischemia

关 键 词：促血管生成素-1 血管内皮细胞 生长因子 基因治疗 大鼠 血管通透性 副作用 

分 类 号：R543[医药卫生—心血管疾病]