Department of Anesthesiology, the Second Affiliated Hospital of Jiaxing University, Nanhu District, No. 1518, Huancheng North Road, Jiaxing, 314033, Zhejiang, China.
Department of Anesthesiology, Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang, China.
Neurotox Res. 2021 Jun;39(3):566-574. doi: 10.1007/s12640-020-00316-8. Epub 2021 Jan 14.
Parkinson's disease (PD) is a severe neurodegenerative disease lacking effective clinical therapies. It is reported that astrocyte-associated neuroinflammation and oxidative stress are involved in the pathological mechanism of PD. In the present study, we aimed to investigate the protective effect of febuxostat against 1 methyl 4 phenyl pyridine (MPP+)-induced injury on primary astrocytes to highlight the potential therapeutic property of febuxostat in PD.MPP+ was used to induce an in vitro PD model in primary rat astrocytes. The levels of ROS and intracellularly reduced GSH were determined using DCFH-DA assay and a commercial GSH kit, respectively. MTT and LDH release assays were utilized to evaluate the cell viability of astrocytes. The expressions of IL-8, IL-1β, TNF-α, MMP-2, and MMP-9 in the astrocytes were detected using qRT-PCR and ELISA assays. QRT-PCR and Western blot analysis were used to determine the expression levels of GFAP in astrocytes. The expression of p-JNK and nuclear levels of NF-κB p65 were evaluated using Western blot analysis. The transcriptional activity of NF-κB was measured using the luciferase activity assay.Firstly, the elevated levels of ROS and decreased levels of intracellularly reduced GSH in primary astrocytes induced by MPP+ were significantly ameliorated by febuxostat. Secondly, treatment with febuxostat rescued MPP+-induced reduction in cell viability and increased LDH release. Thirdly, febuxostat alleviated MPP+-induced inflammatory responses in astrocytes by reducing the expressions of IL-8, IL-1β, TNF-α, GFAP, MMP-2, and MMP-9. Importantly, we found that febuxostat mitigated activation of the JNK/NF-κB signaling pathway by inhibiting the phosphorylation of JNK and nuclear translocation of NF-κB p65.Febuxostat attenuated MPP+-induced inflammatory response by suppressing the JNK/NF-κB signaling pathway in astrocytes.
帕金森病（PD）是一种严重的神经退行性疾病，缺乏有效的临床治疗方法。有报道称，星形胶质细胞相关的神经炎症和氧化应激参与了 PD 的病理机制。在本研究中，我们旨在研究别嘌醇醇对 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPP+）诱导的原代星形胶质细胞损伤的保护作用，以突出别嘌醇醇在 PD 中的潜在治疗特性。
用 MPP+诱导原代大鼠星形胶质细胞体外 PD 模型。使用 DCFH-DA 测定法和商业 GSH 试剂盒分别测定 ROS 和细胞内还原型 GSH 的水平。MTT 和 LDH 释放测定法用于评估星形胶质细胞的细胞活力。使用 qRT-PCR 和 ELISA 测定法检测星形胶质细胞中 IL-8、IL-1β、TNF-α、MMP-2 和 MMP-9 的表达。qRT-PCR 和 Western blot 分析用于确定星形胶质细胞中 GFAP 的表达水平。使用 Western blot 分析评估 JNK 的磷酸化和核内 NF-κB p65 的水平。使用荧光素酶活性测定法测量 NF-κB 的转录活性。
首先，MPP+诱导的原代星形胶质细胞中 ROS 水平升高和细胞内还原型 GSH 水平降低，用别嘌醇醇治疗后明显改善。其次，别嘌醇醇治疗可挽救 MPP+诱导的细胞活力降低和 LDH 释放增加。第三，别嘌醇醇通过降低 IL-8、IL-1β、TNF-α、GFAP、MMP-2 和 MMP-9 的表达，减轻 MPP+诱导的星形胶质细胞炎症反应。重要的是，我们发现别嘌醇醇通过抑制 JNK 的磷酸化和 NF-κB p65 的核转位，减轻了 JNK/NF-κB 信号通路的激活。
别嘌醇醇通过抑制 JNK/NF-κB 信号通路减轻 MPP+诱导的星形胶质细胞炎症反应。
