Early glycolytic reprogramming controls microglial inflammatory activation.
Early glycolytic reprogramming controls microglial inflammatory activation.
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DOI：
10.1186/s12974-021-02187-y
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发表时间：
2021-06-09
影响因子：
9.3
通讯作者：
Zheng LT
中科院分区：
文献类型：
--
作者：
Cheng J;Zhang R;Xu Z;Ke Y;Sun R;Yang H;Zhang X;Zhen X;Zheng LT
小胶质细胞激活介导的神经炎症在神经退行性疾病的进展中起重要作用。小胶质细胞的炎症激活通常伴随着从氧化磷酸化到有氧糖酵解的代谢转换。然而，糖酵解在小胶质细胞活化和神经炎症中的作用和分子机制尚不完全清楚。采用酶联免疫吸附试验（ELISA）、定量逆转录聚合酶链反应（RT-PCR）、Western blot、免疫沉淀、流式细胞术、核因子κB （NF-κB）荧光素酶报告基因检测等方法，对脂多糖（LPS）激活的BV-2小胶质细胞或原代小胶质细胞进行体外糖酵解抑制的抗炎作用及其机制进行研究。在1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）或脂多糖诱导的帕金森病（PD）模型中，通过免疫荧光染色、行为测试和Western blot分析，检测糖酵解抑制剂2-脱氧葡萄糖（2- dg）在体内的抗炎和神经保护作用。我们发现，LPS可迅速增加小胶质细胞的糖酵解，糖酵解抑制剂（2- dg和3-溴丙酮酸（3-BPA））、siRNA葡萄糖转运蛋白1型（Glut-1）和siRNA己糖激酶（HK） 2可消除LPS诱导的小胶质细胞活化。机制研究表明，糖酵解抑制剂显著抑制lps诱导的雷帕霉素机制靶点（mTOR）、核因子-κB激酶亚基β (IKKβ)抑制剂和NF-κB抑制剂α (i -κB -α)磷酸化、i -κB α降解、NF-κB p65亚基核易位和NF-κB转录活性。此外，2-DG显著抑制lps诱导的赖氨酸310上p65/RelA的乙酰化，该乙酰化是由nad依赖性蛋白去乙酰化酶sirtuin-1 （SIRT1）介导的，对NF-κB激活至关重要。一项共培养研究表明，2-DG降低了活化的小胶质细胞对MES23.5多巴胺能神经元细胞的细胞毒性，但没有直接的保护作用。在lps诱导的PD模型中，2-DG显著改善了神经炎症和随后的酪氨酸羟化酶（TH）阳性细胞损失。此外，在mptp诱导的PD模型中，2-DG还能降低多巴胺能细胞死亡和小胶质细胞活化。总的来说，我们的结果表明糖酵解积极参与小胶质细胞的激活。抑制糖酵解可改善与小胶质细胞激活相关的神经炎性疾病。在线版本包含补充材料，可在10.1186/s12974-021-02187-y获得。
Microglial activation-mediated neuroinflammation plays an important role in the progression of neurodegenerative diseases. Inflammatory activation of microglial cells is often accompanied by a metabolic switch from oxidative phosphorylation to aerobic glycolysis. However, the roles and molecular mechanisms of glycolysis in microglial activation and neuroinflammation are not yet fully understood. The anti-inflammatory effects and its underlying mechanisms of glycolytic inhibition in vitro were examined in lipopolysaccharide (LPS) activated BV-2 microglial cells or primary microglial cells by enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot, immunoprecipitation, flow cytometry, and nuclear factor kappa B (NF-κB) luciferase reporter assays. The anti-inflammatory and neuroprotective effects of glycolytic inhibitor, 2-deoxoy-d-glucose (2-DG) in vivo were measured in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-or LPS-induced Parkinson’s disease (PD) models by immunofluorescence staining, behavior tests, and Western blot analysis. We found that LPS rapidly increased glycolysis in microglial cells, and glycolysis inhibitors (2-DG and 3-bromopyruvic acid (3-BPA)), siRNA glucose transporter type 1 (Glut-1), and siRNA hexokinase (HK) 2 abolished LPS-induced microglial cell activation. Mechanistic studies demonstrated that glycolysis inhibitors significantly inhibited LPS-induced phosphorylation of mechanistic target of rapamycin (mTOR), an inhibitor of nuclear factor-kappa B kinase subunit beta (IKKβ), and NF-kappa-B inhibitor alpha (IκB-α), degradation of IκBα, nuclear translocation of p65 subunit of NF-κB, and NF-κB transcriptional activity. In addition, 2-DG significantly inhibited LPS-induced acetylation of p65/RelA on lysine 310, which is mediated by NAD-dependent protein deacetylase sirtuin-1 (SIRT1) and is critical for NF-κB activation. A coculture study revealed that 2-DG reduced the cytotoxicity of activated microglia toward MES23.5 dopaminergic neuron cells with no direct protective effect. In an LPS-induced PD model, 2-DG significantly ameliorated neuroinflammation and subsequent tyrosine hydroxylase (TH)-positive cell loss. Furthermore, 2-DG also reduced dopaminergic cell death and microglial activation in the MPTP-induced PD model. Collectively, our results suggest that glycolysis is actively involved in microglial activation. Inhibition of glycolysis can ameliorate microglial activation-related neuroinflammatory diseases. The online version contains supplementary material available at 10.1186/s12974-021-02187-y.
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DOI：
10.1158/1541-7786.mcr-15-0213
发表时间：
2015-12
期刊：
Molecular cancer research : MCR
影响因子：
--
作者：
Gao Y;Gartenhaus RB;Lapidus RG;Hussain A;Zhang Y;Wang X;Dan HC
通讯作者：
Dan HC
影响因子：
64.5
作者：
Chang CH;Curtis JD;Maggi LB Jr;Faubert B;Villarino AV;O'Sullivan D;Huang SC;van der Windt GJ;Blagih J;Qiu J;Weber JD;Pearce EJ;Jones RG;Pearce EL
通讯作者：
Pearce EL
影响因子：
4.8
作者：
Chen, J;Zhou, YG;Li, G
通讯作者：
Li, G
DOI：
10.1146/annurev.pathol.4.110807.092250
发表时间：
2010
期刊：
Annual review of pathology
影响因子：
--
作者：
Haigis MC;Sinclair DA
通讯作者：
Sinclair DA
影响因子：
4.8
作者：
Kiernan, R;Brès, V;Benkirane, M
通讯作者：
Benkirane, M