FTY720 Inhibits MPP -Induced Microglial Activation by Affecting NLRP3 Inflammasome Activation
FTY720 Inhibits MPP -Induced Microglial Activation by Affecting NLRP3 Inflammasome Activation
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FTY720 通过影响 NLRP3 炎性体激活来抑制 MPP 诱导的小胶质细胞激活
DOI：
10.1007/s11481-019-09843-4
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发表时间：
2019
影响因子：
6.2
通讯作者：
Zhou Hong
中科院分区：
文献类型：
--
作者：
Yao Shu;Li Longjun;Sun Xin;Hua Jun;Zhan Keqi;Hao Li;Liu Lixin;Shi Dongyan;Zhou Hong
帕金森病(PD)是以黑质致密部(SNPC)多巴胺能神经元变性和小胶质细胞过度激活为特征的疾病。本研究旨在观察1-磷酸鞘氨醇受体拮抗剂Fingolimod(FTY720)对1-甲基-4-苯基-1，2，3，6-四氢吡啶(MPTP)诱导的帕金森病动物模型的治疗作用，并探讨其作用机制。C57BL/6J小鼠在皮下注射MPTP前先灌胃FTY720。采用旷场试验和旋转棒试验确定FTY720的疗效。用免疫组织化学、高效液相色谱法和流式细胞仪检测多巴胺能神经元的损伤和单胺类神经递质的产生。免疫荧光(CD68阳性)和酶联免疫吸附试验分析小胶质细胞的激活情况，Western blotting检测激活的信号分子水平。我们的研究结果表明，FTY720显著减轻MPTP诱导的行为缺陷，减少多巴胺能神经元的丢失，并增加多巴胺的释放。FTY720直接抑制MPTP诱导的SNPC小胶质细胞活化，抑制1-甲基-4-苯基吡啶处理的BV-2小胶质细胞产生IL-6、IL-1β和肿瘤坏死因子-α，从而减少SH-SY5Y神经母细胞瘤细胞的凋亡。此外，在MPP+处理的BV-2细胞和原代小胶质细胞中，FTY720显著抑制PI3K/AKT/GSK-3β的磷酸化，减少ROS的产生和P65的激活，并抑制NLRP3炎症体和Caspase-1的激活。结论：FTY720可能通过影响小胶质细胞中ROS的生成和P65的激活来抑制NLRP3炎性小体的激活，从而延缓PD的进展。FTY720可能通过抑制PI3K/AKT/GSK-3β信号通路减少ROS的产生，同时减少p65的磷酸化，从而通过这两条途径减少NLRP3炎症体的激活，最终减轻小胶质细胞激活引起的神经元损伤。
Parkinson’s disease (PD) is characterized by the degeneration of dopaminergic neurons and excessive microglial activation in the substantia nigrapars compacta(SNpc). In the present study, we aimed to demonstrate the therapeutic effectiveness of the potent sphingosine-1-phosphate receptor antagonist fingolimod (FTY720) in an animal model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and to identify the potential mechanisms underlying these therapeutic effects. C57BL/6J mice were orally administered FTY720 before subcutaneous injection of MPTP. Open-field and rotarod tests were performed to determine the therapeutic effect of FTY720. The damage to dopaminergic neurons and the production of monoamine neurotransmitters were assessed using immunohistochemistry, high-performance liquid chromatography, and flow cytometry. Immunofluorescence (CD68- positive) and enzyme-linked immunosorbent assay were used to analyze the activation of microglia, and the levels of activated signaling molecules were measured using Western blotting. Our findings indicated that FTY720 significantly attenuated MPTP-induced behavioral deficits, reduced the loss of dopaminergic neurons, and increased dopamine release. FTY720 directly inhibited MPTP-induced microglial activation in the SNpc, suppressed the production of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in BV-2 microglial cells treated with 1-methyl-4-phenylpyridinium (MPP+), and subsequently decreased apoptosis in SH-SY5Y neuroblastoma cells. Moreover, in MPP+-treated BV-2 cells and primary microglia, FTY720 treatment significantly attenuated the increases in the phosphorylation of PI3K/AKT/GSK-3β, reduced ROS generation and p65 activation, and also inhibited the activation of NLRP3 inflammasome and caspase-1. In conclusion, FTY720 may reduce PD progression by inhibiting NLRP3 inflammasome activation via its effects on ROS generation and p65 activation in microglia. These findings provide novel insights into the mechanisms underlying the therapeutic effects of FTY720, suggesting its potential as a novel therapeutic strategy against PD.Graphical AbstractFTY720 may reduce ROS production by inhibiting the PI3K/AKT/GSK-3β signaling pathway, while at the same time reducing p65 phosphorylation, thus decreasing NLRP3 inflammasome activation through these two pathways, ultimately reducing microglia activation-induced neuronal damage.