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

解剖学报 ›› 2024, Vol. 55 ›› Issue (2) : 133-142.

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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

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*
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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*
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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。

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.

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

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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段兆达 杨力 陈浩伦 刘腾腾 郑立扬 徐冬垚 吴春云. 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[J]. 解剖学报. 2024, 55(2): 133-142 https://doi.org/10.16098/j.issn.0529-1356.2024.02.002
DUAN Zhao-da YANG Li CHEN Hao-lun LIU Teng-teng ZHENG Li-yang XU Dong-yao WU Chun-yun. Scutellarin inhibitting BV-2 microglia-mediated neuroinflammation via the cyclic GMP-AMP synthase - stimulator of interferon gene pathway[J]. Acta Anatomica Sinica. 2024, 55(2): 133-142 https://doi.org/10.16098/j.issn.0529-1356.2024.02.002
中图分类号:      R363.21   

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基金

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

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