NRG1转染对骨髓基质干细胞表达VEGF影响.docVIP

NRG1转染对骨髓基质干细胞表达VEGF影响 　　[摘要]目的 探讨神经调节因子1（neuregulin-1，NRG1）转染是否可以促进骨髓基质干细胞（bone marrow stromal cells，BMSCs）表达血管内皮生长因子（vascular endothelial growth factor，VEGF）。方法 提取与培养3只SD大鼠的BMSCs，采用NRG1质粒转染BMSCs并提取上清液，采用ELISA法测定转染组上清液中VEGF的含量，将非转染BMSCs的样本作为对照组，采用免疫组化方法检测细胞VEGF的表达。结果 BMSCs多数细胞处于伸展状态，呈梭形；经ELISA方法检测，转染组上清液VEGF的含量随时间增加呈增长趋势，与对照组比较，差异有统计学意义（P0.05）；经免疫组化检测，转染组细胞VEGF阳性率为91.07%，对照组细胞阳性率为70.82 %，差异有统计学意义（P0.05）。结论 NRG1转染可以促进BMSCs表达VEGF。 　　[关键词]骨髓基质干细胞；细胞培养；血管内皮生长因子；神经调节因子1 　　[中图分类号] R329 [文献标识码] A [文章编号] 1674-4721（2017）11（b）-0004-04 　　[Abstract]Objective To explore whether bone marrow stromal stem cells （BMSCs） could been promoted by NRG1 transfection to VEGF expression.Methods The BMSCs of three SD rats were isolated and cultured，BMSCs were transfected with NRG1 plasmids，and the supernates were extracted.The contents of VEGF in the supernates were detected by the method of ELISA，and the samples of BMSCs were regarded as the control group.The expression of VEGF in BMSCs was detected by immunohistochemical method.Results Most of BMSCs were stretched and spindle-shaped.The levels of VEGF in the transfection group were increased with time by ELISA detection，compared with the control group，there was significant difference （P0.05）.The VEGF positve cell rate of the transfection group was 91.07% by immunohistochemical detection，while the positive cell rate of the control group was 70.82%，there was significant difference compared with the control group （P0.05）.Conclusion BMSCs can promoted by NRG1 transfection to express VEGF. 　　[Key words]Bone marrow stromal cells；Cell culture；Vascular endothelial growth factor；Neuregulin-1 　　骨髓基?|干细胞（bone marrow stromal stem cells，BMSCs）因其低免疫源性、稳定的扩增能力和无伦理问题等优势被应用于治疗缺血性疾病的研究[1]，然而移植后BMSCs存活率过低的问题一直未被解决[2]。神经调节因子1可通过激活血管内皮ErbB2、ErbB3和ErbB4受体促进血管新生，从而改善细胞存活的微环境[3]。血管内皮生长因子是血管内皮细胞特异性的肝素结合生长因子，可促进内皮细胞生长及血管新生[4]。有报道显示，NRG1与VEGF的表达相关联。而NRG1对BMSCs表达VEGF的影响，未被查见相关报道[5]。本研究将NRG1基因转染到BMSCs，检测其VEGF的表达，旨在为提高移植后BMSCs存活率的相关研究奠定理论基础，现报道如下。 　　1材料与方法