中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (4): 677-683.doi: 10.4103/1673-5374.230294

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

Rho激酶抑制剂Y27632和法舒地尔促进脊髓小胶质细胞迁移的途径

  

  • 收稿日期:2018-02-05 出版日期:2018-04-15 发布日期:2018-04-15
  • 基金资助:

    国家自然科学基金（81471200, 81771341）

The Rho-associated kinase inhibitors Y27632 and fasudil promote microglial migration in the spinal cord via the ERK signaling pathway

Pei-Cai Fu1, Rong-Hua Tang1, Zhi-Yuan Yu1, 2, Min-Jie Xie1, 2, Wei Wang1, 2, Xiang Luo1   

  1. 1 Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province,China
    2 Key Laboratory of Neurological Diseases (Huazhong University of Science and Technology), Ministry of Education of China, Wuhan, Hubei Province, China
  • Received:2018-02-05 Online:2018-04-15 Published:2018-04-15
  • Contact: Xiang Luo, M.D.,tjhfile@tjh.tjmu.edu.cn
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81471200, 81771341.

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摘要：

ROCK又称Rho激酶，已被证实参与中枢神经系统主要免疫细胞小胶质细胞的炎性分泌。课题组以往研究发现，ROCK抑制剂可作用于ERK信号通路增强小胶质细胞的摄取活性，但其对小胶质细胞迁移是否可有影响尚不明确。为观察ROCK抑制剂Y27632和法舒地尔对脊髓原代小胶质细胞迁移功能的影响及其作用机制，实验对脊髓小胶质细胞进行原代培养后，采用Y27632、法舒地尔及ERK抑制剂U0126干预处理，应用免疫荧光标记观察脊髓原代小胶质细胞形态变化，Transwell小室检测小胶质细胞迁移功能，通过In-cell Western Blot检测磷酸化的ERK的表达。结果显示：(1) Y27632和法舒地尔干预可促进小胶质细胞迁移，并使小胶质细胞形态不规则，伸出多个长的枝杈，上调ERK磷酸化蛋白表达；(2) ERK抑制剂U0126干预可抑制Y27632和法舒地尔的上述作用。从而说明，ROCK抑制剂Y27632和法舒地尔应该是通过ERK信号途径，促进脊髓小胶质细胞迁移的。

orcid:0000-0001-9233-5788(Xiang Luo)

关键词: 神经再生, 脊髓损伤, 小胶质细胞, ROCK, Y27632, 法舒地尔, 迁移, 细胞形态, ERK, U0126, In-cell western blot, Transwell小室

Abstract:

Rho-associated kinase (ROCK) is a key regulatory protein involved in inflammatory secretion in microglia in the central nervous system. Our previous studies showed that ROCK inhibition enhances phagocytic activity in microglia through the extracellular signal-regulated kinase (ERK) signaling pathway, but its effect on microglial migration was unknown. Therefore, in this study, we investigated the effects of the ROCK inhibitors Y27632 and fasudil on the migratory activity of primary cultured microglia isolated from the spinal cord, and we examined the underlying mechanisms. The microglia were treated with Y27632, fasudil and/or the ERK inhibitor U0126. Cellular morphology was observed by immunofluorescence. Transwell chambers were used to assess cell migration. ERK levels were measured by incell western blot assay. Y27632 and fasudil increased microglial migration, and the microglia were irregularly shaped and had many small processes. These inhibitors also upregulated the levels of phosphorylated ERK protein. The ERK inhibitor U0126 suppressed these effects of Y27632 and fasudil. These findings suggest that the ROCK inhibitors Y27632 and fasudil promote microglial migration in the spinal cord through the ERK signaling pathway.