人重组血管内皮生长因子-A截短形式的异源表达及其在伤口愈合中的生物学活性。
Heterologous expression of a truncated form of human recombinant vascular endothelial growth factor-A and its biological activity in wound healing.
作者信息
Khaki Mohsen, Salmanian Ali Hatef, Mosayebi Ghasem, Baazm Maryam, Babaei Saeed, Molaee Neda, Abtahi Hamid
机构信息
Molecular and Medicine Research Center, Department of Immunology and Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
出版信息
Iran J Basic Med Sci. 2017 Jul;20(7):791-797. doi: 10.22038/IJBMS.2017.9011.
OBJECTIVES
Vascular endothelial growth factor (VEGF) is one of the most effective proteins in angiogenesis, mesenchymal stem cells (MSCs) differentiation and wound healing. These abilities are therapeutic potential of VEGF in diabetic retinopathy, nephropathy and other tissue damage circumstances. In this study, recombinant VEGF was produced in () system and then biological activity of this protein was evaluated in animal wound healing.
MATERIALS AND METHODS
BL21 (DE3) competent cells were transformed with pET32a-VEGF clone and induced by isopropyl-β-D-thio-galactoside (IPTG). The recombinant protein was purified by affinity chromatography. Recombinant VEGF-A-based ointment (VEGF/Vaseline 0.8 mg/100 w/w) was used for external wound (25×15mm thickness) healing in animal model. activity of ointment was evaluated by clinical evidences and cytological microscopic assessment.
RESULTS
The recombinant protein with molecular weight of 45 kilodaltons (kDa) and concentration of 0.8 mg/ml was produced. Immunoblotting data showed that the antigenic region of VEGF can be expressed in and the recombinant protein has similar epitopes with close antigenic properties to the natural form. Macroscopic findings and microscopic data showed that the recombinant VEGF-A ointment was effective on excisional wound healing.
CONCLUSION
Recombinant VEGF-A produced by pET32a in , possesses acceptable structure and has wound healing capability.
目的
血管内皮生长因子（VEGF）是血管生成、间充质干细胞（MSCs）分化和伤口愈合中最有效的蛋白质之一。这些能力是VEGF在糖尿病视网膜病变、肾病和其他组织损伤情况下的治疗潜力。在本研究中，重组VEGF在（）系统中产生，然后在动物伤口愈合中评估该蛋白质的生物活性。
材料与方法
用pET32a-VEGF克隆转化BL21（DE3）感受态细胞，并用异丙基-β-D-硫代半乳糖苷（IPTG）诱导。重组蛋白通过亲和层析纯化。基于重组VEGF-A的软膏（VEGF/凡士林0.8mg/100w/w）用于动物模型中外部伤口（25×15mm厚度）的愈合。通过临床证据和细胞学显微镜评估来评估软膏的活性。
结果
产生了分子量为45千道尔顿（kDa）、浓度为0.8mg/ml的重组蛋白。免疫印迹数据表明，VEGF的抗原区域可以在（）中表达，并且重组蛋白具有与天然形式相似的表位，抗原特性相近。宏观观察结果和显微镜数据表明，重组VEGF-A软膏对切除伤口的愈合有效。
结论
pET32a在（）中产生的重组VEGF-A具有可接受的结构并具有伤口愈合能力。
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