Fluoxetine suppresses inflammatory reaction in microglia under OGD/R challenge via modulation of NF-kappa B signaling
Fluoxetine suppresses inflammatory reaction in microglia under OGD/R challenge via modulation of NF-kappa B signaling
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氟西汀通过调节 NF-kappa B 信号传导抑制 OGD/R 刺激下小胶质细胞的炎症反应
DOI：
10.1042/bsr20181584
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发表时间：
2019
影响因子：
4
通讯作者：
Hongbin Yuan
中科院分区：
文献类型：
--
作者：
Mouli Tian;Mei Yang;Zhenjie Li;Yiru Wang;Wei Chen;Liye Yang;Yonghua Li;Hongbin Yuan
我们的目的是研究氟西汀（一种选择性血清素再摄取抑制剂）在小胶质细胞（MG）中的抗炎作用以及氧糖剥夺/复氧（OGD/R）下的机制。 BV-2 细胞的 OGD/R 模型用于研究缺血性中风中缺血/再灌注损伤下的小胶质细胞。应用慢病毒转染来敲低 IκB-α。采用酶联免疫吸附法（ELISA）检测TNF-α、IL-1β、IL-6的水平，采用实时荧光定量PCR（real-time PCR）检测IκB-α蛋白的表达。应用蛋白质印迹法分析 NF-κB 信号相关蛋白，并使用细胞计数试剂盒-8 (CCK-8) 评估细胞活力。采用分子对接和药物亲和响应靶稳定性（DARTS）测定法检测IκB-α与氟西汀之间的相互作用。我们发现氟西汀降低了 OGD/R 下 BV-2 细胞上清液中 TNF-α、IL-1β 和 IL-6 以及 NF-κB 亚基 p65 和 p50 的水平。氟西汀通过抑制 IκB-α 泛素化显着提高 IκB-α 水平，并促进 OGD/R 下 BV-2 细胞中 IκB-α 与氟西汀的结合。敲低 IκB-α 减弱了氟西汀导致 OGD/R 下 BV-2 细胞中 TNF-α、IL-1β、IL-6 以及 p65 和 p50 的降低作用。总之，我们目前的研究证明了氟西汀的抗炎作用及其与缺血/再灌注挑战下 MG 中 NF-κB 相关信号调节相关的机制。
We aimed to investigate the anti-inflammatory role of fluoxetine, a selective serotonin reuptake inhibitor, in microglia (MG) and the mechanisms under oxygen glucose deprivation/reoxygenation (OGD/R). An OGD/R model on BV-2 cells was used for the study of microglia under ischemia/reperfusion injury in ischemic stroke. Lentiviral transfection was applied to knock down IκB-α. Enzyme-linked immunosorbent assay (ELISA) was used for detecting levels of TNF-α, IL-1β, and IL-6, and real-time PCR was used to assess the expression of IκB-α protein. Western blotting was applied to analyze NF-κB-signaling related proteins and Cell Counting Kit-8 (CCK-8) was used for assessing cell viability. Molecular docking and drug affinity responsive target stability (DARTS) assay were used for the detection of the interaction between IκB-α and fluoxetine. We found that fluoxetine decreased the levels of TNF-α, IL-1β, and IL-6 in supernatant as well as NF-κB subunits p65 and p50 in BV-2 cells under OGD/R. Fluoxetine significantly increased the level of IκB-α through the inhibition of IκB-α ubiquitylation and promoted the bonding of IκB-α and fluoxetine in BV-2 cells under OGD/R. Knocking down IκB-α attenuated the decreasing effect of TNF-α, IL-1β, and IL-6 as well as p65 and p50 in BV-2 cells under OGD/R led to by fluoxetine. In conclusion, our present study demonstrated the anti-inflammatory role of fluoxetine and its mechanisms related to the modulation of NF-κB-related signaling in MG under ischemia/reperfusion challenge.