血管生成素-1可改善糖尿病伤口中内皮祖细胞依赖性新生血管形成。
Angiopoietin-1 improves endothelial progenitor cell-dependent neovascularization in diabetic wounds.
作者信息
Balaji Swathi, Han Nate, Moles Chad, Shaaban Aimen F, Bollyky Paul L, Crombleholme Timothy M, Keswani Sundeep G
机构信息
Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH.
Division of Infectious Diseases, Department of Medicine, Stanford University School of Medicine, Stanford, CA.
出版信息
Surgery. 2015 Sep;158(3):846-56. doi: 10.1016/j.surg.2015.06.034.
BACKGROUND
The diabetic phenotype of wound healing is in part characterized by impaired neovascularization and deficient endothelial progenitor cell (EPC) recruitment. Angiopoietin-1 (Ang-1) is a potent mobilizer of EPCs from the bone marrow (BM). A suggested mechanism for EPC mobilization from the BM is mediated by matrix metalloproteinase 9 (MMP-9) and stem cell factor (SCF). Taken together, we hypothesized that overexpression of Ang-1 in diabetic wounds will recruit EPCs and improve neovascularization and wound healing.
METHODS
An endothelial lineage BM-labeled murine model of diabetes was developed to track BM-derived EPCs. FVBN mice were lethally irradiated and then reconstituted with BM from syngeneic Tie2/LacZ donor mice. Diabetes was induced with streptozotocin. Dorsal wounds in BM-transplanted mice were treated with Ad-Ang-1, Ad-GFP, or phosphate-buffered saline. At day 7 after injury, wounds were harvested and analyzed. A similar experiment was conducted in EPC mobilization deficient MMP-9 -/- mice to determine whether the effects of Ang-1 were EPC-dependent.
RESULTS
Overexpression of Ang-1 resulted in greatly improved re-epithelialization, neovascularization, and EPC recruitment in diabetic BM-transplanted wounds at day 7. Ang-1 treatment resulted in increased serum levels of proMMP-9 and SCF but had no effect on vascular endothelial growth factor levels. According to our FACS results, peripheral blood EPC (CD34(+)/Cd133(+)/Flk1(+)) counts at day 3 after wounding showed impaired EPC mobilization in MMP-9 -/- mice compared with those of wild-type controls. EPC mobilization was rescued by SCF administration, validating this model for EPC-mobilization-deficient mechanistic studies. In MMP-9 -/- mice, Ad-Ang-1 accelerated re-epithelialization in a similar manner, but had no effect on neovascularization.
CONCLUSION
Our results show that Ang-1 administration results in improved neovascularization which is dependent on EPC recruitment and has direct effects on wound re-epithelialization. These data may represent a novel strategy to correct the phenotype of impaired diabetic neovascularization and may improve diabetic wound healing.
背景
伤口愈合的糖尿病表型部分特征为新生血管形成受损和内皮祖细胞（EPC）募集不足。血管生成素-1（Ang-1）是一种从骨髓（BM）中有效动员EPC的物质。一种从BM动员EPC的推测机制是由基质金属蛋白酶9（MMP-9）和干细胞因子（SCF）介导的。综上所述，我们假设在糖尿病伤口中过表达Ang-1将募集EPC并改善新生血管形成和伤口愈合。
方法
建立一种内皮谱系BM标记的糖尿病小鼠模型以追踪BM来源的EPC。对FVBN小鼠进行致死性照射，然后用同基因Tie2/LacZ供体小鼠的BM进行重建。用链脲佐菌素诱导糖尿病。对BM移植小鼠的背部伤口用Ad-Ang-1、Ad-GFP或磷酸盐缓冲盐水进行处理。在损伤后第7天，收集伤口并进行分析。在EPC动员缺陷的MMP-9 -/-小鼠中进行了类似实验，以确定Ang-1的作用是否依赖于EPC。
结果
在第7天，Ang-1的过表达导致糖尿病BM移植伤口的再上皮化、新生血管形成和EPC募集得到显著改善。Ang-1治疗导致血清中前MMP-9和SCF水平升高，但对血管内皮生长因子水平无影响。根据我们的流式细胞术结果，与野生型对照相比，在受伤后第3天，MMP-9 -/-小鼠外周血EPC（CD34(+)/Cd133(+)/Flk1(+)）计数显示EPC动员受损。通过给予SCF可挽救EPC动员，验证了该模型用于EPC动员缺陷机制研究的有效性。在MMP-9 -/-小鼠中，Ad-Ang-1以类似方式加速了再上皮化，但对新生血管形成无影响。
结论
我们的结果表明，给予Ang-1可改善新生血管形成，这依赖于EPC募集，并对伤口再上皮化有直接影响。这些数据可能代表一种纠正糖尿病新生血管形成受损表型的新策略，并可能改善糖尿病伤口愈合。
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