Reactive Astrocytes Display Pro-inflammatory Adaptability with Modulation of Notch-PI3K-AKT Signaling Pathway Under Inflammatory Stimulation
Reactive Astrocytes Display Pro-inflammatory Adaptability with Modulation of Notch-PI3K-AKT Signaling Pathway Under Inflammatory Stimulation
复制标题
反应性星形胶质细胞在炎症刺激下通过调节 Notch-PI3K-AKT 信号通路表现出促炎适应性
DOI：
10.1016/j.neuroscience.2020.05.023
复制
发表时间：
2020-08-01
期刊：
影响因子：
3.3
通讯作者：
Chen, Bei-Yu
中科院分区：
文献类型：
--
作者：
Cheng, Ying-Ying;Ding, Yin-Xiu;Chen, Bei-Yu
星形胶质细胞是辅助中枢神经系统（CNS）功能的主要胶质细胞，但反应性星形胶质细胞在中枢神经系统疾病中的功能变化和调节机制仍然知之甚少。本研究培养小鼠原代星形胶质细胞，并进行炎症损伤，通过免疫荧光、PCR、Western blot、CCK-8和抑制实验观察星形胶质细胞的功能变化以及Notch-PI3K-AKT信号激活的参与情况。在星形胶质细胞中清楚地观察到Notch下游信号Hes-1，Notch信号抑制剂GSI剂量依赖性地降低了切割的Notch-1水平，而不影响细胞活力。脂多糖加干扰素γ（LPS+IFNγ）的炎症损伤诱导活化星形胶质细胞中蛋白质和mRNA水平的促炎细胞因子（即iNOS、IL-1β、IL-6和TNF）增加，而GSI治疗可减少或阻断这种增加。星形胶质细胞的细胞活力在不同组之间没有表现出显着差异。虽然证实了 MyD88、NF-κ B 和磷-NF-κ B 的增加，但在 LPS+IFN γ 损伤的活化星形胶质细胞中观察到 PI3K、AKT 和磷-AKT 的上调，并通过 GSI 治疗而减少。抑制剂实验表明，抑制 Notch-PI3K-AKT 信号激活可减少 LPS + IFN γ 炎症损伤引发的促炎细胞因子的产生。这项研究表明，反应性星形胶质细胞通过响应炎症刺激而激活Notch-PI3K-AKT信号传导而表现出促炎适应性，这表明反应性星形胶质细胞中的Notch-PI3K-AKT通路可能作为对抗中枢神经系统炎症性疾病的有希望的靶标。 (C) 2020 国际广播组织。由爱思唯尔有限公司出版。保留所有权利。
Astrocytes are major glial cells critical in assisting the function of the central nervous system (CNS), but the functional changes and regulation mechanism of reactive astrocytes are still poorly understood in CNS diseases. In this study, mouse primary astrocytes were cultured, and inflammatory insult was performed to observe functional changes in astrocytes and the involvement of Notch-PI3K-AKT signaling activation through immunofluorescence, PCR, Western blot, CCK-8, and inhibition experiments. Notch downstream signal Hes-1 was clearly observed in the astrocytes, and Notch signal inhibitor GSI dose-dependently decreased the cleaved Notch-l level without an influence on cell viability. Inflammatory insult of lipopolysaccharide plus interferon-gamma (LPS+IFN gamma) induced an increase in pro-inflammatory cytokines, that is, iNOS, IL-1 beta, IL-6, and TNF, at the protein and mRNA levels in activated astrocytes, which was reduced or blocked by GSI treatment. The cell viability of the astrocytes did not show significant differences among different groups. While an increase in MyD88, NF-kappa B, and phosphor-NF-kappa B was confirmed, upregulation of PI3K, AKT, and phosphor-AKT was observed in the activated astrocytes with LPS+IFN gamma insult and was reduced by GSI treatment. Inhibitor experiments showed that inhibition of Notch-PI3K-AKT signaling activation reduced the pro-inflammatory cytokine production triggered by LPS +IFN gamma inflammatory insult. This study showed that the reactive astrocytes displayed pro-inflammatory adaptability through Notch-PI3K-AKT signaling activation in response to inflammatory stimulation, suggesting that the Notch-PI3K-AKT pathway in reactive astrocytes may serve as a promising target against CNS inflammatory disorders. (C) 2020 IBRO. Published by Elsevier Ltd. All rights reserved.