Diao Yongpeng, Lian Lishan, Guo Lilong, Chen Houzao, Chen Yuexin, Song Xiaojun, Li Yongjun
Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
Chin Med J (Engl). 2014;127(20):3623-9.
Therapeutic angiogenesis has been shown to promote blood vessel growth and improve tissue perfusion. Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis. However, it has side effects that limit its therapeutic utility in vivo, especially at high concentrations. This study aimed to investigate whether an intramuscular injection of a genetically engineered zinc finger VEGF-activating transcription factor modulates the endothelial progenitor cells (EPC) and promotes therapeutic angiogenesis in a hindlimb ischemia model with type 1 diabetes.
Alloxan (intravenous injection) was used to induce type I diabetes in C57BL/6 mice (n = 58). The ischemic limb received ZFP-VEGF (125 µg ZFP-VEGF plasmid in 1% poloxamer) or placebo (1% poloxamer) intramuscularly. Mice were sacrificed 3, 5, 10, or 20 days post-injection. Limb blood flow was monitored using laser Doppler perfusion imaging. VEGF mRNA and protein expression were examined using real-time PCR and ELISA, respectively. Capillary density, proliferation, and apoptosis were examined using immunohistochemistry techniques. Flow cytometry was used to detect the EPC population in bone marrow. Two-tailed Student's paired t test and repeated-measures analysis of variance were used for statistical analysis.
ZFP-VEGF increased VEGF mRNA and protein expression at 3 and 10 days post-injection, and increased EPC in bone marrow at day 5 and 20 post-injection compared with controls (P < 0.05). ZFP-VEGF treatment resulted in better perfusion recovery, a higher capillary density and proliferation, and less apoptosis compared with controls (P < 0.05).
Intramuscular ZFP-VEGF injection promotes therapeutic angiogenesis in an ischemic hindlimb model with type 1 diabetes. This might be due to the effects of VEGF on cell survival and EPC recruitment.
治疗性血管生成已被证明可促进血管生长并改善组织灌注。血管内皮生长因子（VEGF）在血管生成中起重要作用。然而，它具有副作用，限制了其在体内的治疗效用，尤其是在高浓度时。本研究旨在调查在1型糖尿病后肢缺血模型中，肌肉注射基因工程锌指VEGF激活转录因子是否能调节内皮祖细胞（EPC）并促进治疗性血管生成。
用四氧嘧啶（静脉注射）诱导C57BL/6小鼠（n = 58）患1型糖尿病。缺血肢体肌肉注射ZFP-VEGF（125μg ZFP-VEGF质粒溶于1%泊洛沙姆）或安慰剂（1%泊洛沙姆）。注射后3、5、10或20天处死小鼠。使用激光多普勒灌注成像监测肢体血流。分别用实时PCR和ELISA检测VEGF mRNA和蛋白表达。用免疫组织化学技术检测毛细血管密度、增殖和凋亡。用流式细胞术检测骨髓中的EPC群体。采用双尾学生配对t检验和重复测量方差分析进行统计学分析。
与对照组相比，ZFP-VEGF在注射后3天和10天增加了VEGF mRNA和蛋白表达，在注射后5天和20天增加了骨髓中的EPC（P < 0.05）。与对照组相比，ZFP-VEGF治疗导致更好的灌注恢复、更高的毛细血管密度和增殖以及更少的凋亡（P < 0.05）。
肌肉注射ZFP-VEGF可促进1型糖尿病缺血后肢模型中的治疗性血管生成。这可能是由于VEGF对细胞存活和EPC募集的作用。
