中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8411-8415.doi: 10.3969/j.issn.1673-8225.2010.45.013

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

短时低频电刺激体外培养许旺细胞髓鞘的形成　

万丽丹1，丁文龙2，李  峰2，夏  蓉2，朱  浩2，刘德明1，林雪群1   

  1. 1南昌大学医学院解剖学教研室，江西省南昌市 330006；2上海交通大学医学院解剖学教研室，上海市  200025
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 作者简介:万丽丹☆，女，1980年生，江西省南昌市人，汉族，2009年上海交通大学医学院毕业，博士，讲师，主要从事神经损伤与再生修复方面的研究。 wanld1980@gmail.com
  • 基金资助:

    上海市重点学科建设项目(S30201)，南昌大学博士科研基金。　

Effect of short-term low-frequency electrical stimulation on myelin formation of Schwann cells in vitro

Wan Li-dan1, Ding Wen-long2, Li Feng2, Xia Rong2, Zhu Hao2, Liu De-ming1, Lin Xue-qun1   

  1. 1 Department of Anatomy, Medical College of Nanchang University, Nanchang  330006, Jiangxi Province, China; 2 Department of Anatomy, Shanghai Jiao Tong University, School of Medicine Shanghai  200025, China
  • Online:2010-11-05 Published:2010-11-05
  • About author:Wan Li-dan☆, Doctor, Lecturer, Department of Anatomy, Medical College of Nanchang University, Nanchang 330006, Jiangxi Province, China wanld1980@gmail.com
  • Supported by:

     the Shanghai Leading Academic Discipline Project, No. S30201*; the Doctoral Scientific Research Foundation of Nanchang University*

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摘要：

背景：周围神经系统损伤后，短时低频电刺激已被证明可显著促进轴突再生和选择性功能修复，但目前对电刺激是否影响周围神经髓鞘的形成还知之甚少，而电刺激发挥作用究竟是通过神经元还是许旺细胞还有待证实。
目的：建立体外背根神经元与许旺细胞联合培养模型，观察短时低频电刺激对许旺细胞髓鞘形成的影响。
方法：体外培养背根神经元，纯化后预先施予电刺激(20 Hz，100 μs，3 V)，持续作用1 h，24 h后再加入许旺细胞悬液制成背根神经元/许旺细胞联合培养模型。在此基础上，用L-ascorbic acid诱导髓鞘形成，分别于诱导后第7，14天观察培养体系中髓鞘的形成。
结果与结论：电刺激增强背根神经元分泌脑源性神经营养因子(P < 0.05)，经电刺激作用的背根神经元再与许旺细胞联合培养，最终表现为髓鞘形成增多以及髓鞘蛋白表达上调(P < 0.05)。提示短时低频电刺激对体外许旺细胞髓鞘的形成具有促进作用，初步认为该作用至少通过刺激神经元分泌脑源性神经营养因子增多导致。

关键词: 电刺激, 髓鞘, 联合培养, 背根神经元, 许旺细胞, 神经组织构建

Abstract:

BACKGROUND: Short-time low-frequency electrical stimulation has the potential to promote axonal regeneration and target selection after peripheral nerve injury. However, studies concerning the effect of electrical stimulation on myelination are still elusive, and the possible target cell (neuron or Schwann cells) on which electrical stimulation impacts remains unknown.
OBJECTIVE: To establish the co-culture model of dorsal root ganglion neurons and Schwann cells, and investigate the short-time low-frequency electrical stimulation effect on Schwann cell myelination in vitro.
METHODS: Purified dorsal root ganglion neurons were subjected to continuous electrical stimulation (20 Hz, 100 μs, 3 V) for one hour. 24 hours later, the purified Schwann cells suspension was added into pre-stimulated neurons/Schwann cell cultures. At 7 and 14 days after L-ascorbic acid induction, myelin formation was observed and determined.
RESULTS AND CONCLUSION: Electrical stimulation enhanced brain-derived neurotrophic factor secretion from purified dorsal root neurons (P < 0.05). Dorsal root neurons treated with electrical stimulation cocultured with Schwann cells, which presented increased myelination and upregulated myelin protein expression (P < 0.05). These results have indicated that short-term low-frequency electrical stimulation promoted Schwann cell myelination in vitro, and the effect was at least initially mediated via the enhancement of neuronal brain-derived neurotrophic factor signals.

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