首页|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

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

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

ScutellarinBV-2 microgliaCyclic GMP-AMP synthetase-stimulator of interferon gene pathwayPYD domains-containing protein 3NeuroinflammationImmunofluorescenceWestern blotting

段兆达、杨力、陈浩伦、刘腾腾、郑立扬、徐冬垚、吴春云

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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

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

国家自然科学基金云南省教育厅科研项目研究生项目

819602232023Y0793

2024

解剖学报
中国解剖学会

解剖学报

CSTPCD
影响因子:0.462
ISSN:0529-1356
年,卷(期):2024.55(2)
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