中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (9): 1483-1490.doi: 10.4103/1673-5374.165521

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

促红细胞生成素基因修饰神经干细胞移植修复损伤脊髓效果更好

  

  • 收稿日期:2015-08-17 出版日期:2015-09-28 发布日期:2015-09-28
  • 基金资助:

    吉林省科技发展计划资助项目(2011084)

Transplantation of erythropoietin gene-modified neural stem cells improves the repair of injured spinal cord

Min-fei Wu1, Shu-quan Zhang2, Rui Gu3, Jia-bei Liu3, Ye Li3, Qing-san Zhu3   

  1. 1 Department of Orthopedics, the Second Hospital of Jilin University, Changchun, Jilin Province, China
    2 Department of Orthopedics, Tianjin Nankai Hospital, Tianjin, China
    3 Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun, Jilin Province, China
  • Received:2015-08-17 Online:2015-09-28 Published:2015-09-28
  • Contact: Rui Gu, M.D., fandiliucl@126.com.
  • Supported by:

    This study was supported by the Science and Technology Development Program of Jilin Province of China, No. 2011084. 

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摘要：

促红细胞生成素对脊髓损伤保护作用的研究仍不深入。为此，实验将真核表达质粒pcDNA3.1人源促红细胞生成素转染至体外培养的大鼠神经干细胞中，按照自由落体打击法建立脊髓损伤大鼠模型，将转染后的神经干细胞注入其蛛网膜下腔为促红细胞生成素-神经干细胞组，蛛网膜下腔注入未转染神经干细胞的脊髓损伤大鼠作为神经干细胞组，注入DMEM/F12培养液的脊髓损伤大鼠作为脊髓损伤组。损伤后1-4周，大鼠下肢运动功能从好到差依次为：促红细胞生成素-神经干细胞组、神经干细胞组、脊髓损伤组。损伤后72 h，与脊髓损伤组相比，促红细胞生成素-神经干细胞组脊髓损伤区周围细胞凋亡指数、Caspase-3基因和蛋白表达显著降低，bcl-2基因和蛋白表达显著升高。损伤后4周，脊髓损伤区周围脊髓空洞、运动和感觉诱发电位潜伏期：两个干细胞治疗组明显低于脊髓损伤组，尤以促红细胞生成素-神经干细胞组作用最明显。CM-Dil阳性细胞和HRP阳性神经纤维数、运动和感觉诱发电位波幅：仍以两个干细胞治疗组明显高于脊髓损伤组，尤以促红细胞生成素-神经干细胞组作用最明显。结果证实，人源促红细胞生成素基因修饰神经干细胞蛛网膜下腔移植可用于修复脊髓损伤，促进脊髓功能恢复，其效果较未转染促红细胞生成素的神经干细胞移植更好，实验结果有转化医学借鉴意义。

关键词: 神经再生, 脊髓损伤, 神经干细, 促红细胞生成素, 神经再生, 运动功能, 大鼠, 蛛网膜下腔, 移植, 损伤, 恢复

Abstract:

The protective effects of erythropoietin on spinal cord injury have not been well described. Here, the eukaryotic expression plasmid pcDNA3.1 human erythropoietin was transfected into rat neural stem cells cultured in vitro. A rat model of spinal cord injury was established using a free falling object. In the human erythropoietin-neural stem cells group, transfected neural stem cells were injected into the rat subarachnoid cavity, while the neural stem cells group was injected with non-transfected neural stem cells. Dulbecco’s modified Eagle’s medium/F12 medium was injected into the rats in the spinal cord injury group as a control. At 1–4 weeks post injury, the motor function in the rat lower limbs was best in the human erythropoietin-neural stem cells group, followed by the neural stem cells group, and lastly the spinal cord injury group. At 72 hours, compared with the spinal cord injury group, the apoptotic index and Caspase-3 gene and protein expressions were apparently decreased, and the bcl-2 gene and protein expressions were noticeably increased, in the tissues surrounding the injured region in the human erythropoietin-neural stem cells group. At 4 weeks, the cavities were clearly smaller and the motor and somatosensory evoked potential latencies were remarkably shorter in the human erythropoietin-neural stem cells group and neural stem cells group than those in the spinal cord injury  group. These differences were particularly obvious in the human erythropoietin-neural stem cells group. More CM-Dil-positive cells and horseradish peroxidase-positive nerve fibers and larger amplitude motor and somatosensory evoked potentials were found in the human erythropoietin-neural stem cells group and neural stem cells group than in the spinal cord injury group. Again, these differences were particularly obvious in the human erythropoietin-neural stem cells group. These data indicate that transplantation of erythropoietin gene-modified neural stem cells into the subarachnoid cavity to help repair spinal cord injury and promote the recovery of spinal cord function better than neural stem cell transplantation alone. These findings may lead to significant improvements in the clinical treatment of spinal cord injuries.

Key words: erve regeneration, spinal cord injury, neural stem cells, erythropoietin, motor function, subarachnoid cavity, transplantation, injury, recovery, neural regeneration