Nurr1通过抑制小胶质细胞NF-κB发挥抗炎作用
Nurr1 attenuates microglia inflammation by inhibiting NF-κB
王伟 1侯晓霖 1陈秋元 2潘晓玥 2彭涛 2王俊燕 2李云鸿 2王银2
作者信息
- 1. 宁夏医科大学总医院,宁夏 银川 750004
- 2. 宁夏医科大学基础医学院,宁夏 银川 750004
- 折叠
摘要
目的 探索在细菌脂多糖(LPS)诱导的小鼠小胶质细胞(BV2 细胞株)炎症反应中,核受体相关蛋白1(Nurr1)是否通过抑制NF-κB的表达及核转位发挥抗炎作用.方法 培养小鼠BV2 小胶质细胞株,将细胞分为Ctrl组、LPS组、C-DIM12(Nurrl特异性激动剂)组、siRNA干扰组、LPS+C-DIM12 组.利用CCK-8 检测LPS和C-DIM12 的最适作用浓度,用Western blot观察NF-κB和Nurr1 的表达变化,用酶联免疫吸附法(ELISA)检测培养液中炎症因子IL-6、IL-1β和TNF-α的水平,用免疫荧光观察siRNA干扰或C-DIM12 激活Nurr1 表达活性后,NF-κB及p-IκB 在 BV2 细胞中的表达水平.结果 LPS 刺激 BV2 细胞后,NF-κB 表达提高,炎症因子IL-1、IL-6、TNF-α释放量增加(P<0.05).C-DIM12 激活Nurr1 后,免疫荧光染色技术和Western blot实验结果都显示NF-κB的表达显著下降,而小干扰RNA(siNurr1)沉默Nurr1 表达后,NF-κB表达增加.在LPS激活的小胶质细胞中,NF-κB二聚体分子的结合分子p-IκB的磷酸化蛋白水平相对于对照组显著升高,而C-DIM12激活Nurr1 后,p-IκB水平显著下降.结论 Nurr1 可以通过抑制小胶质细胞NF-κB表达水平并阻止NF-κB核位移发挥抗炎作用.Nurr1 可能是一个新的潜在的治疗中枢神经系统炎症相关疾病的靶点.
Abstract
Objective To explore whether Nurr1 inhibits NF-κB expression and nuclear translocation in BV2 microglia inflam-matory response induced by LPS to play an anti-inflammatory role.Methods Mouse BV2 microglia cells were cultured and divided in-to Ctrl group,LPS group,C-DIM12 group(Nurr1 specific agonist),siRNA interference group and LPS+C-DIM12 group.CCK-8 was used to detect the optimal concentration of LPS and C-DIM12,and the expression changes of NF-κB and Nurr1 were measured by Western blot,and the inflammatory factors IL-6,IL-1β and TNF-α levels were detected by ELISA.After the expression activity of Nurr1 was disrupted by siRNA or activated by C-DIM12,the expression levels of NF-κB and p-IκB in BV2 cells were investigated.Results After the BV2 cells were stimulated by LPS,the expression of NF-κB increased and the release of inflammatory factors IL-1,IL-6,TNF-α also increased(P<0.05).After Nurr1 was activated by C-DIM12,the expression of NF-κB decreased signif-icantly.However,after small interfering RNA(siNurr1)silenced the expression of Nurr1,the expression of NF-κB increased.In LPS-activated microglia,the level of phosphorylated protein I-κB,which was NF-κB dimer molecule binding molecule,was significantly higher than that in the control group.After Nurr1 was activated by C-DIM12,pI-κB level decreased significantly.Conclusion Nurr1 can inhibit the expression of NF-κB,reverse NF-κB nuclear translocation and protect the CNS from inflammation.Nurr1 may be a new and potential target for the treatment of inflammation related CNS diseases.
关键词
Nurr1/NF-κB/神经炎症/小胶质细胞/p-IκBKey words
Nurr1/NF-κB/Neuroinflammation/Microglia/p-IκB引用本文复制引用
基金项目
国家自然科学基金(82060223)
宁夏回族自治区自然科学基金(2022AAC03607)
宁夏回族自治区自然科学基金(2021AAC03372)
宁夏回族自治区重点研发基金(2021BEG03097)
出版年
2024
NETL
NSTL
万方数据