中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (5): 913-920.doi: 10.4103/1673-5374.249240

• 原著:神经损伤修复保护与再生 • 上一篇    

(5R)-5-羟基雷公藤内酯醇抑制星形胶质细胞中级联反应而产生的炎症

  

  • 出版日期:2019-05-15 发布日期:2019-05-15
  • 基金资助:

    中国国家自然科学基金项目(81402932)

(5R)-5-hydroxytriptolide inhibits the inflammatory cascade reaction in astrocytes

Yan-Qiu Cui 1, 2 , Yan Zheng 3 , Gui-Lian Tan 4 , Dong-Mei Zhang 4 , Jun-Ya Wang 4 , Xiao-Min Wang  2, 5   

  1. 1 Functional Laboratory of Experiment Teaching Center for Basic Medical Sciences, Capital Medical University, Beijing, China
    2 Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Capital Medical University, Beijing, China
    3 Department of Physiology, Capital Medical University, Beijing, China
    4 Department of Basic Medicine, Yanjing Medical College, Capital Medical University, Beijing, China
    5 Beijing Institute for Brain Disorders, Beijing, China
  • Online:2019-05-15 Published:2019-05-15
  • Contact: Xiao-Min Wang, MD, PhD, xmwang@ccmu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81402932

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摘要：

有研究表明，(5R)-5-羟基雷公藤内酯醇是雷公藤内酯的修饰类似物之一，具有比雷公藤内酯更低的细胞毒性和更强的免疫抑制活性，但其是否对神经炎症有预防作用目前尚不清楚。实验设计以20，100，500 nM的(5R)-5-羟基雷公藤内酯醇预处理新生SD大鼠(北京维通利华实验动物技术有限公司)的全脑制备的原代星形胶质细胞1h，然后以1.0 μg/mL脂多糖刺激24h的建立体外神经炎症模型。以Griess试剂检测一氧化氮的产生，以免疫组化染色检测星形胶质细胞标志物胶质纤维酸性蛋白的阳性反应，以ELISA测定培养上清液中的肿瘤坏死因子α和白细胞介素1β的水平，以免疫荧光染色检测NF-κB/p65的表达，以Western blot分析核因子IκB-α磷酸化和NF-κB/P65的位置。结果发现，(5R)-5-羟基雷公藤内酯醇抑制由脂多糖诱导原代星形胶质细胞产生的一氧化氮，降低胶质纤维酸性蛋白的阳性反应强度，减少了培养上清液中肿瘤坏死因子α和白细胞介素1β的表达，抑制了IκB-α的磷酸化以及NF-κB/P65向细胞核的转位，显示了(5R)-5-羟基雷公藤内酯醇对脂多糖诱导的神经胶质炎症反应有抑制作用，为(5R)-5-羟基雷公藤内酯醇治疗神经退行性疾病提供了细胞学实验数据。

orcid: 0000-0001-5346-3859 (Yan-Qiu Cui)

关键词: (5R)-5-羟基雷公藤内酯醇, 神经炎症, 肿瘤坏死因子α, 白细胞介素1β, 一氧化氮, NF-κB/ P65, IκB-α, 神经胶质细胞, 神经再生

Abstract:

Many studies have shown that (5R)-5-hydroxytriptolide is the optimal modified analogue of triptolide, possessing comparable im¬munosuppressive activity but much lower cytotoxicity than triptolide. Whether (5R)-5-hydroxytriptolide has preventive effects on neuroinflammation is unclear. This study was designed to pretreat primary astrocytes from the brains of neonatal Sprague-Dawley rats with 20, 100 and 500 nM (5R)-5-hydroxytriptolide for 1 hour before establishing an in vitro neuroinflammation model with 1.0 μg/mL lipopolysaccharide for 24 hours. The generation of nitric oxide was detected by Griess reagents. Astrocyte marker glial fibrillary acidic pro¬tein was measured by immunohistochemical staining. The levels of tumor necrosis factor-α and interleukin-1β in the culture supernatant were assayed by enzyme linked immunosorbent assay. Nuclear factor-κB/p65 expression was examined by immunofluorescence staining. The phosphorylation of inhibitor of nuclear factor IκB-α and the location of nuclear factor-κB/P65 were determined using western blot assay. Our data revealed that (5R)-5-hydroxytriptolide inhibited the generation of nitric oxide, tumor necrosis factor-α and interleukin-1β from primary astrocytes activated by lipopolysaccharide, decreased the positive reaction intensity of glial fibrillary acidic protein, reduced the expression of tumor necrosis factor alpha and interleukin-1β in culture supernatant, inhibited the phosphorylation of IκB-α and the translocation of nuclear factor-κB/P65 to the nucleus. These results have confirmed that (5R)-5-hydroxytriptolide inhibits lipopolysac¬charide-induced glial inflammatory response and provides cytological experimental data for (5R)-5-hydroxytriptolide in the treatment of neurodegenerative diseases.

Key words: neuroinflammation, (5R)-5-hydroxytriptolide, tumor necrosis factor-α, interleukin-1β, nitric oxide, nuclear factor-κB/P65, IκB-α, microglia, neural regeneration