解剖学报

解剖学报 ›› 2024, Vol. 55 ›› Issue (2): 133-142.doi: 10.16098/j.issn.0529-1356.2024.02.002

• 神经生物学 • 上一篇    下一篇

灯盏乙素通过环状GMP-AMP合酶-干扰素基因刺激因子通路抑制BV-2小胶质细胞介导的神经炎症

段兆达 杨力 陈浩伦 刘腾腾 郑立扬 徐冬垚 吴春云*   

  1. 昆明医科大学基础医学院人体解剖学与组织学胚胎学系，昆明 650500
  • 收稿日期:2023-03-23 修回日期:2023-07-03 出版日期:2024-04-06 发布日期:2024-04-06
  • 通讯作者: 吴春云 E-mail:wuchunyunkm@163.com
  • 基金资助:
    国家自然科学基金;云南省教育厅科学研究基金研究生项目

Scutellarin inhibitting BV-2 microglia-mediated neuroinflammation via the cyclic GMP-AMP synthase - stimulator of interferon gene pathway

DUAN  Zhao-da YANG  Li  CHEN  Hao-lun LIU  Teng-teng  ZHENG  Li-yang XU  Dong-yao  WU  Chun-yun*   

  1. Department of Human Anatomy and Histology and Embryology, School of Basic Medical Sciences, Kunming Medical University, Kunming650500, China
  • Received:2023-03-23 Revised:2023-07-03 Online:2024-04-06 Published:2024-04-06
  • Contact: WU Chun-yun E-mail:wuchunyunkm@163.com

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摘要：

目的 探讨灯盏乙素对脂多糖（LPS）诱导的BV-2小胶质细胞神经炎症的影响。  方法 培养BV-2小胶质细胞系，将BV-2小胶质细胞分为对照组（Ctrl）、环状GMP-AMP合酶(cGAS)抑制剂RU320521（RU.521）组、LPS组、LPS+RU.521组、LPS+灯盏乙素预处理(LPS+S)组、LPS+S+RU.521组，共6组。Western blotting及免疫荧光双标染色法检测并观察BV-2小胶质细胞中cGAS、干扰素基因刺激因子(STING)、核因子κB（NF-κB）、磷酸化NF-κB（p-NF-κB)、PYD结构域蛋白3 (NLRP3)和肿瘤坏死因子α（TNF-α）的表达变化（n=3）。  结果 Western blotting和免疫荧光双标染色均显示，与对照组相比，LPS诱导后，BV-2小胶质细胞中cGAS、STING、p-NF-κB、NLRP3和TNF-α蛋白的表达水平显著升高（P＜0.05）；与LPS组相比，LPS+S组中cGAS、STING、p-NF-κB、NLRP3和TNF-α蛋白的表达水平显著下降（P＜0.05）。使用cGAS通路抑制剂RU.521后显示了与灯盏乙素预处理组相似的作用效果。此外，NFκB在各组的变化不明显（P＞0.05）。  结论 灯盏乙素干预抑制BV-2小胶质细胞介导的神经炎症反应，可能与cGAS-STING 信号通路有关。

关键词: 灯盏乙素, BV-2小胶质细胞, 环状GMP-AMP合酶-干扰素基因刺激因子通路, PYD结构域蛋白3, 神经炎症, 免疫荧光, 免疫印迹法

Abstract:

Objective To explore the effect of scutellarin on lipopolysaccharide (LPS) induced neuroinflammation in BV-2 microglia cells.   Methods  BV-2 microglia were cultured and randomly divided into 6 groups: control group (Ctrl), cyclic GMP-AMP synthetase（cGAS）inhibitor RU320521 group (RU.521 group), LPS group, LPS+RU.521 group, LPS+ scutellarin pretreatment group (LPS+S) and LPS+S+RU.521 group. The expressions of cGAS, stimulator of interferon gene (STING), nuclear factor kappa B (NF-κB), phosphorylated NF-κB (p-NF-κB), neuroinflammatory factors PYD domainscontaining protein 3（NLRP3) and tumor necrosis factor α（TNF-α）in BV-2 microglia were detected by Western blotting and immunofluorescent double staining (n=3).   Results Western blotting and immunofluorescent double staining showed that compared with the control group, the expression of cGAS, STING, p-NF-κB, NLRP3 and TNF-α in BV-2 microglia increased significantly after LPS induction(P＜0.05), while the expression of cGAS, STING, p-NF-κB, NLRP3 and TNF-α in LPS+S group were significantly lower than those in LPS group (P＜0.05). Treatment with cGAS pathway inhibitor RU.521 showed similar effects as the pre-treatment group with scutellarin. In addition, the change of NF-κB in each group was not statistically significant（P＞0.05）.   Conclusion Scutellarin inhibits the neuroinflammation mediated by BV-2 microglia cells, which may be related to cGAS-STING signaling pathway.

Key words:  Scutellarin, BV-2 microglia, Cyclic GMP-AMP synthetase-stimulator of interferon gene pathway, PYD domains-containing protein3, Neuroinflammation, Immunofluorescence, Western blotting 

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