中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (16): 1465-1472.doi: 10.3969/j.issn.1673-5374.2013.16.003

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

Toll样受体4在海马神经元炎症反应中的作用

  

  • 收稿日期:2012-12-11 修回日期:2013-03-17 出版日期:2013-06-05 发布日期:2013-06-05

Role of Toll-like receptor 4 in inflammatory reactions of hippocampal neurons

Yae Hu1, Jiahui Mao1, Yu Zhang2, Ailing Zhou1   

  1. 1 Department of Pathophysiology, Medical School of Nantong University, Nantong 226001, Jiangsu Province, China
    2 Department of Pediatrics, the First People’s Hospital of Nantong, Nantong 226001, Jiangsu Province, China
  • Received:2012-12-11 Revised:2013-03-17 Online:2013-06-05 Published:2013-06-05
  • Contact: Ailing Zhou, Master, Professor, Department of Pathophysiology, Medical School of Nantong University, Nantong 226001, Jiangsu Province, China, alz@ntu.edu.cn.
  • About author:Yae Hu★, Master, Associate professor.

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摘要：

脂多糖能激活免疫细胞上的Toll样受体4介导免疫反应。然而Toll样受体4在原代培养的海马神经元上是否表达以及在脂多糖刺激海马神经元产生炎症反应中的作用至今尚无共识。实验采用脂多糖刺激大鼠海马神经元，利用Toll样受体4抗体阻断技术分析脂多糖刺激Toll样受体4介导的信号通路及炎性反应变化。免疫荧光检测显示，Toll样受体4在海马神经元胞膜上呈阳性；RT-PCR和Western blot检测显示，脂多糖刺激后Toll样受体4 mRNA和蛋白水平、核转录因子κB p65蛋白表达、转化生长因子α和白细胞介素1β mRNA的表达均增加，且胞浆中IκBα降解明显增加；Toll样受体4的抗体几乎完全阻断上述现象。提示在大鼠海马神经元上脂多糖主要通过Toll样受体4激活核转录因子κB信号通路，参与神经炎症反应，说明神经元不仅是神经炎症反应中的受害者，可能也是神经炎症的执行者。

关键词: 神经再生, Toll样受体4, 脂多糖, 核转录因子κB, 白细胞介素1β, 肿瘤坏死因子α, 海马, 神经元, 大鼠, 炎症, 基金资助文章

Abstract:

Lipopolysaccharide stimulates Toll-like receptor 4 on immune cells to produce immune mediators. Toll-like receptor 4 is also expressed by non-immune cells, which can be stimulated by lipopolysaccharide. However, whether Toll-like receptor 4 is expressed by primary cultured hippocampal neurons and its specific role in lipopolysaccharide-induced neuroinflammation is currently undefined. In this study, Toll-like receptor 4 antibody blocking was used to analyze the Toll-like receptor 4 signaling pathway and changes in inflammation of lipopolysaccharide stimulated hippocampal neurons. Immunofluorescence showed that Toll-like receptor 4 protein was mainly located in the membrane of hippocampal neurons. Quantitative reverse transcription-PCR and western blot assay showed that after stimulation of lipopolysaccharide, the mRNA and protein levels of Toll-like receptor 4 and the mRNA levels of interleukin-1β and tumor necrosis factor-α were significantly increased. In addition, there was increased phosphorylation and degradation of kappa B α inhibitor in the cytosol and increased nuclear factor-κB p65 expression in the nuclei. Pretreatment with Toll-like receptor 4 antibody could almost completely block this increase. These experimental findings indicate that lipopolysaccharide participates in neuroinflammation by stimulating Toll-like receptor 4/nuclear factor-κB pathway in hippocampal neurons, which may be both “passive victims” and “activators” of neuroinflammation.

Key words: neural regeneration, inflammation, Toll-like receptor 4, lipopolysaccharide, nuclear factor-kappa B, interleukin-1 beta, tumor necrosis factor-alpha, hippocampus, neurons, grants-supported paper, neuroregeneration