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 (11): 1860-1864.doi: 10.4103/1673-5374.219048

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

桂枝提取物下调TLR4/MyD88信号通路减轻BV2细胞的神经炎症反应

  

  • 收稿日期:2017-08-26 出版日期:2017-11-15 发布日期:2017-11-15
  • 基金资助:

      国家自然科学基金（81473383）；中国医学科学院医学与健康科技创新工程项目（2016-I2M-3-007）； 国家科技重大专项项目（2012ZX09103101-078 和2017ZX09101003-003-019）

  Ramulus Cinnamomi extract attenuates neuroinflammatory responses via downregulatingTLR4/MyD88 signaling pathway in BV2 cells

Huan Yang1, Xiao Cheng1, Ying-lin Yang1, Yue-hua Wang1,2, Guan-hua Du1, 2   

  1. 1 Beijing Key Laboratory of Drug Target Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
    2 State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical  Sciences & Peking Union Medical College, Beijing, China
  • Received:2017-08-26 Online:2017-11-15 Published:2017-11-15
  • Contact: Yue-hua Wang or Guan-hua Du,wangyuehua@pku.org.cn or dugh@imm.ac.cn.
  • Supported by:

    This study was supported by a grant from the National Natural Science Foundation of China, No.81473383; a grant from the Medical and Health Innovation Project of Chinese Academy of Medical Sciences, No. 2016-I2M-3-007; and a grant from Key Project of New-Drugs Creation of Science and Technology of China, No. 2012ZX09103101-078 and 2017ZX09101003-003-019.

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摘要：

药用植物桂枝已被证实可以缓解炎症反应。为探索桂枝提取物对于脂多糖诱导的小胶质细胞BV2神经炎症反应的作用及相关机制,实验将BV2细胞分别在含有脂多糖，脂多糖+30 μg/mL桂枝提取物，脂多糖+100 μg/mL桂枝提取物的培养基及常规培养基（对照组）进行培养。在光学显微镜下观察BV2细胞形态，运用MTT 方法检测细胞活性。分别用Griess 试剂及ELISA法检测BV2细胞中NO，白细胞介素-1和1β，及肿瘤坏死因子α水平。 Western Blot 法检测BV2细胞中COX-2, TLR4 和 MyD88 蛋白表达。可见30、100 μg/mL桂枝提取物，对脂多糖诱导的BV2细胞活性无明显影响。脂多糖显著增加了BV2细胞中NO，白细胞介素6和1β及肿瘤坏死因子α的水平，而30 和100 μg/mL桂枝提取物明显降低了上述促炎症细胞因子的水平。此外，桂枝提取物显著抑制了在脂多糖诱导的BV2细胞中COX-2, TLR4和MyD88蛋白的表达。结果证实，桂枝提取物可以减轻神经炎症反应，这可能与下调TLR4/MyD88信号通路有关。

orcid:0000-0002-7650-6417(Yue-hua Wang)

关键词: 神经再生, 桂枝, BV2细胞系, 脂多糖, 神经炎症, 炎症介质, TLR4/MyD88信号通路, NO, 白细胞介素6, 白细胞介素1β, 肿瘤坏死因子α

Abstract:

Ramulus Cinnamomi (RC), a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide (LPS)-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium (control group), LPS, LPS plus 30 μg/mL RC extract, or LPS plus 100 μg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay.Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1β, and tumor necrosis factor α in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor α in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating
the TLR4/MyD88 signaling pathway.

Key words: nerve regeneration, Ramulus Cinnamomi, BV2 cells, lipopolysaccharide, neuroinflammation, pro-inflammatory factors, TLR4/MyD88 signaling pathway, nitric oxide, interleukin-6, interleukin-1β, tumor necrosis factor α, neuronal regeneration