低氧预处理脐带间充质干细胞移植促进大鼠脊髓损伤修复

发布时间：2018-08-27 17:54

【摘要】：目的:评估低氧预处理脐带间充质干细胞(human umbilical cord mesenchymal stem cells,UCMSCs)移植修复大鼠急性脊髓损伤的作用并初步探讨其机制。方法:分别于常氧和低氧条件下培养UCMSCs,细胞免疫荧光鉴定细胞,ELISA测定细胞脑源性生长因子(BDNF)、血管源性生长因子(VEGF)、睫状生长因子(CNTF)和肝细胞生长因子(HGF)的浓度。采用改良的Al1en’s法(2.5 g×10 cm)建立60只大鼠T10脊髓损伤模型,随机分为假手术组、模型组、低氧细胞移植组及常氧细胞移植组,每组20只,损伤后即刻移植1×10~6个细胞至低氧细胞移植组及常氧细胞移植组动物脊髓内。术前及术后每周进行运动功能BBB评分;采用免疫荧光法检测移植细胞的存活、神经细胞凋亡;采用ELISA法检测细胞移植第2、14、28 d脊髓组织肝细胞生长因子的浓度。结果:在低氧组,体外由UCMSCs分泌的BDNF、VEGF、CNTF和HGF浓度显著高于常氧组细胞。与模型组比较,细胞移植组运动功能评分和HGF含量增加,而神经细胞的凋亡率降低,但以低氧细胞移植组改变最明显。结论:低氧预处理的UMCSCs具有提高神经营养因子分泌作用,其机制可能与下调神经细胞凋亡,增加受损脊髓组织HGF有关。
[Abstract]:Aim: to evaluate the effect of hypoxic preconditioning (human umbilical cord mesenchymal stem cells,UCMSCs) transplantation on the repair of acute spinal cord injury (sci) in rats. Methods: UCMSCs, cells were cultured under normoxic and hypoxic conditions, respectively. The concentrations of (BDNF), (VEGF), ciliary growth factor (CNTF) and hepatocyte growth factor (HGF) were determined by Elisa. 60 T10 spinal cord injury models were established by modified Al1en's method (2.5 g 脳 10 cm) and were randomly divided into sham-operation group, model group, hypoxic cell transplantation group and normoxic cell transplantation group, with 20 rats in each group. Immediately after injury, 1 脳 10 ~ 6 cells were transplanted into the spinal cord of rats in hypoxic cell transplantation group and normoxic cell transplantation group. BBB score of motor function was performed before and every week after transplantation; survival of transplanted cells and apoptosis of nerve cells were detected by immunofluorescence method; and the concentration of hepatocyte growth factor in spinal cord tissue was detected by ELISA method on the 28th day after transplantation. Results: the concentrations of BDNF,VEGF,CNTF and HGF secreted by UCMSCs in hypoxia group were significantly higher than those in normoxic group. Compared with the model group, the motor function score and HGF content in the transplantation group increased, while the apoptosis rate of the nerve cells decreased, but the changes were most obvious in the hypoxic cell transplantation group. Conclusion: hypoxia preconditioning UMCSCs can increase the secretion of neurotrophic factor, and its mechanism may be related to down-regulation of neuronal apoptosis and increase of HGF in injured spinal cord tissue.
【作者单位】：南方医科大学珠江医院骨科;
【基金】：国家自然科学基金-青年科学基金(81400996) 广东省自然科学基金-博士启动项目(2014A030310100)
【分类号】：R651.2

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