中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (2): 276-282.doi: 10.4103/1673-5374.200811

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

电针对脊髓损伤大鼠RhoA和ROCKⅡ表达的影响

  

  • 收稿日期:2016-12-29 出版日期:2017-02-15 发布日期:2017-02-15
  • 基金资助:

    国家自然科学基金（81360562）

Effect of electroacupuncture on the mRNA and protein expression of Rho-A and Rho-associated kinase II in spinal cord injury rats

You-jiang Min1, Li-li-qiang Ding1, 2, Li-hong Cheng1, Wei-ping Xiao1, Xing-wei He1, Hui Zhang1, Zhi-yun Min1, Jia Pei1   

  1. 1 Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi Province, China; 
    2 Shanghai University of Traditional Chinese Medicine, Shanghai, China
  • Received:2016-12-29 Online:2017-02-15 Published:2017-02-15
  • Contact: Li-hong Cheng or Wei-ping Xiao, M.D., 495466620@qq.com or jxxiaowp@163.com.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81360562.

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摘要：

 

电针可促进损伤脊髓修复，但其具体作用机制仍不清楚。Rho/ROCK信号通路可通过控制细胞的粘附与迁移来调控细胞骨架，抑制中枢神经系统损伤后的轴突再生，从而阻碍脊髓损伤后的修复。为此，我们假设电针能够通过抑制Rho/ROCK信号通路发挥修复脊髓损伤的作用。实验设计了通过使用重物打击法制作脊髓损伤大鼠模型，观察电针腰阳关(GV3)，大椎(GV14)，双侧足三里(ST36)和次髎穴(BL32)和/或单唾液酸神经节苷脂肌肉注射对模型大鼠的干预效果。发现与两种方法联合治疗相比，两种治疗方法单独治疗均促进了大鼠后肢功能恢复。但实时荧光定量PCR和原位杂交及蛋白印迹显示，持续的电针干预抑制了模型大鼠损伤脊髓组织RhoA和ROCK II的mRNA和蛋白的表达，而神经节苷脂单独治疗及两种方法联合治疗却未能达到此抑制效果。结果说明电针可通过特异性抑制损伤脊髓RhoA和ROCKⅡ的mRNA与蛋白的表达，抑制Rho/ROCK信号通路，发挥对损伤脊髓的保护作用，而电针与神经节苷脂联用未见其协同效应。

ORCID:0000-0003-0786-9510(Li-hong Cheng);0000-0002-8664-5437(Wei-ping Xiao)

关键词: 神经再生, 脊髓损伤, 电针, 神经再生, Rho/ROCK信号通路, 单唾液酸神经节苷脂, 运动功能, 细胞骨架, 实时荧光定量PCR, 蛋白印迹, 原位杂交

Abstract:

Electroacupuncture is beneficial for the recovery of spinal cord injury, but the underlying mechanism is unclear. The Rho/Rho-associated kinase (ROCK) signaling pathway regulates the actin cytoskeleton by controlling the adhesive and migratory behaviors of cells that could inhibit neurite regrowth after neural injury and consequently hinder the recovery from spinal cord injury. Therefore, we hypothesized electroacupuncture could affect the Rho/ROCK signaling pathway to promote the recovery of spinal cord injury. In our experiments, the spinal cord injury in adult Sprague-Dawley rats was caused by an impact device. Those rats were subjected to electroacupuncture at Yaoyangguan (GV3), Dazhui (GV14), Zusanli (ST36) and Ciliao (BL32) and/or monosialoganglioside treatment. Behavioral scores revealed that the hindlimb motor functions improved with those treatments. Real-time quantitative polymerase chain reaction, fluorescence in situ hybridization and western blot assay showed that electroacupuncture suppressed the mRNA and protein expression of Rho-A and Rho-associated kinase II (ROCKII) of injured spinal cord. Although monosialoganglioside promoted the recovery of hindlimb motor function, monosialoganglioside did not affect the expression of Rho-A and ROCKII. However, electroacupuncture combined with monosialoganglioside did not further improve the motor function or suppress the expression of Rho-A and ROCKII. Our data suggested that the electroacupuncture could specifically inhibit the activation of the Rho/ROCK signaling pathway thus partially contributing to the repair of injured spinal cord. Monosialoganglioside could promote the motor function but did not suppress expression of RhoA and ROCKII. There was no synergistic effect of electroacupuncture combined with monosialoganglioside.

Key words: nerve regeneration, spinal cord injury, electroacupuncture, Rho/Rho-associated kinase signaling pathway, monosialoganglioside, motor function, cytoskeleton, real-time quantitative polymerase chain reaction, western blot assay, hybridization in situ, neural regeneration