title: A vascular endothelial growth factor activating transcription factor increases the endothelial progenitor cells population and induces therapeutic angiogenesis in a type 1 diabetic mouse with hindlimb ischemia.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
authors.forename: Yongpeng
authors.initials: Y
authors.lastname: Diao
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
authors.forename: Lishan
authors.initials: L
authors.lastname: Lian
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
authors.forename: Lilong
authors.initials: L
authors.lastname: Guo
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
authors.forename: Houzao
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
authors.forename: Yuexin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Song
authors.affiliation: Department of Vascular Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China. Email: yongj_93@hotmail.com.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism*; D066026:Endothelial Progenitor Cells / Q000378:metabolism*; D005434:Flow Cytometry; D006614:Hindlimb / Q000473:pathology*; D007511:Ischemia / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
chemical_list: D042461:Vascular Endothelial Growth Factor A