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转染人神经生长因子和血管内皮生长因子基因增强大鼠骨髓间充质干细胞的血管生成作用。

Enhancement of angiogenic effect of co-transfection human NGF and VEGF genes in rat bone marrow mesenchymal stem cells.

机构信息

Department of Internal Neurology, Second Affiliated Hospital of Zhengzhou University, Henan 453003, PR China.

出版信息

Gene. 2011 Oct 10;485(2):167-71. doi: 10.1016/j.gene.2011.06.027. Epub 2011 Jul 18.

Abstract

The current study explored the feasibility and efficacy of co-transfection of the human nerve growth factor (NGF) and vascular endothelial growth factor 165 (VEGF165) genes in rat bone marrow mesenchymal stem cells (BMSCs). The obtained hNGF and vascular endothelial growth factor (VEGF) cDNAs were cloned into the pEGFP-C1 expression vector to construct the recombinant vectors. Co-transfection in rat BMSCs was performed and the expressions of both genes were detected by RT-PCR, Western blot, and enzyme-linked immunospecific assay. The biological activity of recombinant NGF and VEGF proteins was confirmed using the Chick Chorioallantoic Membrane (CAM) assay. NGF and VEGF genes could be expressed successfully in rat BMSCs. The recombinant NGF and VEGF from the rat BMSCs showed a more significant synergetic biological activity compared with single recombinant NGF or VEGF. These findings demonstrate that the co-transfection of hNGF+VEGF genes can enhance the angiogenic effect in vivo.

摘要

本研究探讨了在大鼠骨髓间充质干细胞(BMSCs)中转染人神经生长因子(NGF)和血管内皮生长因子 165(VEGF165)基因的可行性和疗效。将获得的 hNGF 和血管内皮生长因子(VEGF)cDNA 克隆到 pEGFP-C1 表达载体中,构建重组载体。通过转染大鼠 BMSCs,采用 RT-PCR、Western blot 和酶联免疫吸附试验检测两种基因的表达。利用鸡胚尿囊膜(CAM)实验证实重组 NGF 和 VEGF 蛋白的生物学活性。NGF 和 VEGF 基因可在大鼠 BMSCs 中成功表达。与单独的重组 NGF 或 VEGF 相比,来自大鼠 BMSCs 的重组 NGF 和 VEGF 表现出更显著的协同生物学活性。这些发现表明,hNGF+VEGF 基因的共转染可以增强体内的血管生成作用。

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