中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (11): 2011-2019.doi: 10.4103/1673-5374.259624

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

胚胎神经干细胞移植到大鼠受损脊髓可抑制疼痛

  

  • 出版日期:2019-11-15 发布日期:2019-11-15
  • 基金资助:

    FAPESP圣保罗州研究支持基金

Pain inhibition through transplantation of fetal neuronal progenitors into the injured spinal cord in rats

Chary M. Batista 1 , Eric D. Mariano 1 , Camila S. Dale 2 , Alexandre F. Cristante 3 , Luiz R. Britto 4 , Jose P. Otoch 5 , Manoel J. Teixeira 1 , Matthias Morgalla 6 , Guilherme Lepski 6, 7   

  1. 1 Department of Neurology, School of Medicine, University of São Paulo, São Paulo, Brazil
    2 Department of Anatomy, Institute of Biomedical Science, University of São Paulo, São Paulo, Brazil
    3 Department of Orthopedic and Traumatology, School of Medicine, University of São Paulo, São Paulo, Brazil
    4 Department of Physiology and Biophysics, Institute of Biomedical Science, University of São Paulo, São Paulo, Brazil
    5 Department of Surgery, School of Medicine, University of São Paulo, São Paulo, Brazil
    6 Department of Neurosurgery, Eberhard-Karls University, Tuebingen, Germany
    7 Department of Psychiatry, School of Medicine, University of São Paulo, São Paulo, Brazil
  • Online:2019-11-15 Published:2019-11-15
  • Contact: Guilherme Lepski, PhD, g.lepski@hc.fm.usp.br.
  • Supported by:

    This work was supported by FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo, No. 2013/12056-0 (to GL) and No. 2015/02154-0 (to CMB).

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摘要：

细胞移植在治疗脊髓损伤后的神经性疼痛方面很有前景，然而就其神经分化潜力和对抗疼痛的有效性而言，理想的细胞类型仍存在很大争议。为评估胚胎神经干细胞缓解慢性疼痛的能力及对运动恢复的影响。在成年Wistar大鼠脊髓损伤后10d于脊髓内注射培养基（假手术组）或Wistar大鼠E14胚胎的端脑泡或腹侧髓质提取的胚胎神经干细胞。在细胞移植后8周内以von Frey细丝法和热板法评估大鼠痛感，以BBB评分和斜板测试评估大鼠运动能力，结果显示移植后4和5周，端脑泡组和腹侧髓质组的热痛觉过敏均有所改善。此外，8周时端脑泡组和腹侧髓质组的机械异常性疼痛均得到改善。与假手术组相比，在端脑泡组或腹侧髓质组中未见明显的运动功能恢复。体视学分析显示~70％的端脑泡组和腹侧髓质组细胞分化为NeuN阳性神经元，端脑泡组中的脑啡肽能和γ-氨基丁酸能细胞比例较高，腹侧髓质组中的脑啡肽能和5-羟色胺能细胞较多。实验表明，胚胎神经元祖细胞植入受损的脊髓后会分化为不同的神经元亚型，主要是γ-氨基丁酸能、5-羟色胺能和脑啡肽能细胞，从而减轻疼痛。该研究于2016年3月4日获得圣保罗大学医学院动物伦理委员会（033/14）批准。

orcid: 0000-0002-9030-4192(Guilherme Lepski)

关键词: 脊髓损伤, 慢性疼痛, 神经干细胞, 细胞移植, 神经元分化, GABA能神经元, 血清素能神经元, 脑啡肽神经元

Abstract:

Neuropathic pain after spinal cord injury (SCI) is a complex condition that responds poorly to usual treatments. Cell transplantation represents a promising therapy; nevertheless, the ideal cell type in terms of neurogenic potential and effectiveness against pain remains largely controversial. Here, we evaluated the ability of fetal neural stem cells (fNSC) to relieve chronic pain and, secondarily, their effects on motor recovery. Adult Wistar rats with traumatic SCI were treated, 10 days after injury, with intra-spinal injections of culture medium (sham) or fNSCs extracted from telencephalic vesicles (TV group) or the ventral medulla (VM group) of E/14 embryos. Sensory (von Frey filaments and hot plate) and motor (the Basso, Beattie, Bresnahan locomotor rating scale and inclined plane test) assessments were performed during 8 weeks. Thereafter, spinal cords were processed for immunofluorescence and transplanted cells were quantified by stereology. The results showed improvement of thermal hyperalgesia in the TV and VM groups at 4 and 5 weeks after transplantation, respectively. Moreover, mechanical allodynia improved in both the TV and VM groups at 8 weeks. No significant motor recovery was observed in the TV or VM groups compared with sham. Stereological analyses showed that ~70% of TV and VM cells differentiated into NeuN+ neurons, with a high proportion of enkephalinergic and GABAergic cells in the TV group and enkephalinergic and serotoninergic cells in the VM group. Our study suggests that neuronal precursors from TV and VM, once implanted into the injured spinal cord, maturate into different neuronal subtypes, mainly GABAergic, serotoninergic, and enkephalinergic, and all subtypes alleviate pain, despite no significant motor recovery. The study was approved by the Animal Ethics Committee of the Medical School of the University of São Paulo (protocol number 033/14) on March 4, 2016.

Key words: spinal cord injuries, chronic pain, neural stem cells, cell transplantation, neuronal differentiation, GABAergic neuron, serotoninergic neuron, enkephalinergic neuron