Engineered zinc finger-activating vascular endothelial growth factor transcription factor plasmid DNA induces therapeutic angiogenesis in rabbits with hindlimb ischemia
Engineered zinc finger-activating vascular endothelial growth factor transcription factor plasmid DNA induces therapeutic angiogenesis in rabbits with hindlimb ischemia
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DOI：
10.1161/01.cir.0000145139.53840.49
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发表时间：
2004-10-19
期刊：
影响因子：
37.8
通讯作者：
Annex, BH
中科院分区：
文献类型：
--
作者：
Dai, QS;Huang, JH;Annex, BH
背景-治疗性血管生成寻求促进血管生长以改善组织灌注。血管内皮生长因子（VEGF）以多种亚型存在。我们研究了一个工程化的锌指转录因子质粒，旨在激活内源性VEGF基因（ZFP-VEGF）。方法和结果-新西兰白色兔（n=56）进行单侧股动脉结扎和切除。术后第10天，缺血肌肉接受ZFP治疗（500 μ g ZFP-VEGF质粒）或不接受ZFP治疗（β-半乳糖苷酶、空质粒或无质粒）。第1组（n=13）于注射后第3天取血，用实时荧光定量聚合酶链反应（PCR）检测VEGF mRNA，用ELISA法检测VEGF蛋白。第2组（n=13）和第3组（n=10）在注射后11天收获。第2组进行组织学研究，第3组进行组织学和血流变化研究。第4组和第5组（每组n=10）分别在注射后22天和32天收获，并研究血流量的变化。在第1组中，ZFP-VEGF治疗组中VEGF（121）、VEGF（165）、VEGF（189）和蛋白质的VEGF mRNA拷贝数显著高于无ZFP治疗组。在第2组和第3组中，治疗组与未治疗组相比，毛细血管密度和增殖细胞显著更大，凋亡显著更低。ZFP治疗组与非ZFP治疗组相比，缺血肢体与非缺血肢体的血流比率变化显著更大（6.57+/-1.52% vs 3.38+/-0.87%，P
Background-Therapeutic angiogenesis seeks to promote blood vessel growth to improve tissue perfusion. Vascular endothelial growth factor (VEGF) exists in multiple isoforms. We investigated an engineered zinc finger-containing transcription factor plasmid designed to activate the endogenous VEGF gene (ZFP-VEGF).Methods and Results-New Zealand White rabbits (n=56) underwent unilateral femoral artery ligation and excision. At day 10 postoperatively, the ischemic muscle received ZFP treatment (500 mug ZFP-VEGF plasmid) or no ZFP treatment (beta-galactosidase, empty, or no plasmid). Group 1 (n=13) was harvested 3 days after injection to examine VEGF mRNA by real-time polymerase chain reaction and protein by ELISA. Groups 2 (n=13) and 3 (n=10) were harvested 11 days after injection. Group 2 was studied by histology and group 3, by histology and changes in blood flow. Groups 4 and 5 (n=10 each) were harvested 22 and 32 days after injection, respectively, and studied for changes in blood flow. In group 1, VEGF mRNA copy numbers were significantly higher for VEGF(121), VEGF(165), VEGF(189), and protein in the ZFP-VEGF-treatment versus no-ZFP-treatment arms. In groups 2 and 3, capillary density and proliferating cells were significantly greater and apoptosis significantly lower in the treatment versus no-treatment arms. Changes in the blood flow ratio of the ischemic to the nonischemic limb were significantly greater in the treatment versus no-ZFP-treatment groups (6.57+/-1.52% versus 3.38+/-0.87%, P