The State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Shanghai Medical College, Fudan University; Department of Neurology, Jinan Central Hospital Affiliated to Shandong University, Jinan, China.
The State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Shanghai Medical College, Fudan University.
Biochim Biophys Acta Mol Basis Dis. 2020 Jan 1;1866(1):165590. doi: 10.1016/j.bbadis.2019.165590. Epub 2019 Nov 7.
Neuroinflammation, as an important pathological characteristic of Parkinson's disease (PD), is primarily mediated by activated astrocytes and microglia. Neuron-restrictive silencer factor/repressor element 1 (RE1)-silencing transcription factor (NRSF/REST) regulates many genes and signal pathways involved in the inflammatory process in astrocytes. In the present study, we established the GFAP-Cre:NRSF conditional knockout (cKO) mice. The expression of inflammation-associated molecules were measured in primary astrocytes from wild type (WT) and cKO mice after stimulation by 1-Methyl-4-phenylpyridine (MPP), LPS, and conditioned medium (CM) of LPS-treated BV-2 microglial cells. The inflammatory molecule expression in BV-2 microglial cells exposed to conditioned medium of MPP-treated primary astrocytes were also analyzed. Moreover, a subacute regimen of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride (MPTP) was used to establish mouse PD model and the damages to the nigrostriatal pathway were comprehensively evaluated in WT and cKO mice. We found that MPP induced a remarkable increase of NRSF expression in cultured astrocytes. Compared to WT astrocytes, the expression of inflammatory molecules IL-1β, IL-6, COX-2, and iNOS increased dramatically in NRSF deficient astrocytes challenged with CM of LPS-treated BV-2 cells. COX-2 and IL-1β transcripts were significantly elevated in BV-2 microglial cells exposed to CM of MPP-treated NRSF deficient astrocytes compared to WT astrocytes. In cKO mice, the activation of astrocytes and microglial cells was more obvious, and the nigrostriatal dopaminergic system was more heavily injured compared to their WT counterparts after MPTP administration. Our results suggest that reactive NRSF deficient astrocytes orchestrated with microglial cells aggravate the pathophysiological progress in PD.
神经炎症是帕金森病（PD）的重要病理特征，主要由激活的星形胶质细胞和小胶质细胞介导。神经元限制沉默因子/沉默元件 1（RE1）-沉默转录因子（NRSF/REST）调节星形胶质细胞中参与炎症过程的许多基因和信号通路。在本研究中，我们建立了 GFAP-Cre：NRSF 条件性敲除（cKO）小鼠。用 1-甲基-4-苯基吡啶（MPP）、LPS 和 LPS 处理的 BV-2 小胶质细胞条件培养基（CM）刺激野生型（WT）和 cKO 小鼠的原代星形胶质细胞后，测量炎症相关分子的表达。还分析了暴露于 MPP 处理的原代星形胶质细胞条件培养基的 BV-2 小胶质细胞中炎症分子的表达。此外，使用 1-甲基-4-苯基-1,2,3,6-四氢吡啶盐酸盐（MPTP）亚急性方案建立小鼠 PD 模型，并在 WT 和 cKO 小鼠中全面评估黑质纹状体通路的损伤。我们发现 MPP 诱导培养的星形胶质细胞中 NRSF 表达显著增加。与 WT 星形胶质细胞相比，NRSF 缺陷星形胶质细胞在 LPS 处理的 BV-2 细胞 CM 刺激下，炎症分子 IL-1β、IL-6、COX-2 和 iNOS 的表达显著增加。与 WT 星形胶质细胞相比，暴露于 MPP 处理的 NRSF 缺陷星形胶质细胞 CM 的 BV-2 小胶质细胞中 COX-2 和 IL-1β 转录物显著升高。与 WT 对照相比，MPTP 给药后 cKO 小鼠的星形胶质细胞和小胶质细胞激活更明显，黑质纹状体多巴胺能系统损伤更严重。我们的研究结果表明，反应性 NRSF 缺陷星形胶质细胞与小胶质细胞共同作用，加剧了 PD 的病理生理进展。
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