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 (1): 132-139.doi: 10.4103/1673-5374.243718

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

叶酸促进许旺细胞增殖、迁移、分泌神经生长因子而修复促进周围神经损伤？

  

  • 出版日期:2019-01-15 发布日期:2019-01-15
  • 基金资助:

    北京市卫生系统高水平卫生技术人员项目(2013-050；基金负责人：闫家智)

Folic acid contributes to peripheral nerve injury repair by promoting Schwann cell proliferation, migration, and secretion of nerve growth factor

Wei-Bo Kang, Yong-Jie Chen, Du-Yi Lu, Jia-Zhi Yan   

  1. Department of Orthopedic Surgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China
  • Online:2019-01-15 Published:2019-01-15
  • Contact: ia-Zhi Yan, MD, beijingtiantan@163.com.
  • Supported by:

    This study was supported by the High Levels of Health Technical Personnel in Beijing City Health System of China, No. 2013-3-050 (to  JZY).

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摘要：

最近研究表明，周围神经损伤后，腹腔内注射叶酸能改善神经电生理结果及增加轴突数量和密度，其具体机制尚未明确。为此，实验旨在探索叶酸通过影响许旺细胞功能促进周围神经损伤修复。(1)实验体外分离、培养，获得大鼠原代许旺细胞。(2)体外实验显示：用含0，10，50，100，150，200 mg/L叶酸和40 ng/L神经生长因子的培养基与许旺细胞培养后CCK-8检测发现，培养72 h，100 mg/L叶酸组的培养孔光密度值较对照组显著增加；培养96 h，50，100，150，200 mg/L叶酸和40 ng/L神经生长因子组的培养孔光密度值较对照组显著增加，100 mg/L叶酸组的培养孔光密度值较50 mg/L叶酸组及40 ng/L神经生长因子组显著增加；在Transwell检测中，100，150 mg/L叶酸组透过滤膜的许旺细胞数显著高于对照组，100 mg/L叶酸组差异更为显著。在NGF-ELISA检测中，培养第3天时，50，100，150 mg/L叶酸组神经生长因子浓度显著高于对照组，而100 mg/L叶酸组神经生长因子浓度显著高于50，150 mg/L叶酸组；培养第7天，10，50，100 mg/L叶酸组神经生长因子浓度显著高于对照组，而50 mg/L叶酸组神经生长因子浓度显著高于10，100 mg/L叶酸组。(3)体内实验显示：大鼠坐骨神经损伤后连续7 d腹腔内注射叶酸80μg/kg，免疫组织化学染色结果显示叶酸组坐骨神经组织许旺细胞数量显著多于单纯坐骨神经损伤大鼠；(4)上述数据说明：叶酸可能通过促进许旺细胞增殖、迁移、分泌神经生长因子，而发挥改善周围神经损伤的修复作用。

orcid: 0000-0001-7404-4688(Jia-Zhi Yan)

关键词: 叶酸, 许旺细胞, 细胞增殖, 细胞迁移, 神经生长因子, 周围神经损伤, 周围神经修复, 神经营养因子, 组织工程, 神经再生

Abstract:

After peripheral nerve injury, intraperitoneal injection of folic acid improves axon quantity, increases axon density and improves electromy¬ography results. However, the mechanisms for this remain unclear. This study explored whether folic acid promotes peripheral nerve injury repair by affecting Schwann cell function. Primary Schwann cells were obtained from rats by in vitro separation and culture. Cell prolif¬eration, assayed using the Cell Counting Kit-8 assay, was higher in cells cultured for 72 hours with 100 mg/L folic acid compared with the control group. Cell proliferation was also higher in the 50, 100, 150, and 200 mg/L folic acid groups compared with the control group after culture for 96 hours. Proliferation was markedly higher in the 100 mg/L folic acid group compared with the 50 mg/L folic acid group and the 40 ng/L nerve growth factor group. In Transwell assays, the number of migrated Schwann cells dramatically increased after culture with 100 and 150 mg/L folic acid compared with the control group. In nerve growth factor enzyme-linked immunosorbent assays, treatment of Schwa nn cell cultures with 50, 100, and 150 mg/L folic acid increased levels of nerve growth factor in the culture medium compared with the con¬trol group at 3 days. The nerve growth factor concentration of Schwann cell cultures treated with 100 mg/L folic acid group was remarkably higher than that in the 50 and 150 mg/L folic acid groups at 3 days. Nerve growth factor concentration in the 10, 50, and 100 mg/L folic acid groups was higher than that in the control group at 7 days. The nerve growth factor concentration in the 50 mg/L folic acid group was re¬markably higher than that in the 10 and 100 mg/L folic acid groups at 7 days. In vivo, 80 μg/kg folic acid was intraperitoneally administrated for 7 consecutive days after sciatic nerve injury. Immunohistochemical staining showed that the number of Schwann cells in the folic acid group was greater than that in the control group. We suggest that folic acid may play a role in improving the repair of peripheral nerve injury by promoting the proliferation and migration of Schwann cells and the secretion of nerve growth factors.

Key words: nerve regeneration, folic acid, Schwann cell, cell functions, peripheral nerve injury, peripheral nerve repair, neurotrophic factor, tissue engineering, neural regeneration, biomaterial, neural regeneration