Division of Regulatory Glycobiology, Tohoku Medical and Pharmaceutical University, Institute of Molecular Biomembrane and Glycobiology, 4-4-1 Komatsushima, Aoba-ku, Sendai, Miyagi 981-8558, Japan; Department of Pharmacology, Pharmacy College, Nantong University, Nantong, Jiangsu Province 226001, China.
Division of Regulatory Glycobiology, Tohoku Medical and Pharmaceutical University, Institute of Molecular Biomembrane and Glycobiology, 4-4-1 Komatsushima, Aoba-ku, Sendai, Miyagi 981-8558, Japan; Clinical Medical Research Center, The Second Affiliated Hospital of Nantong University, Nantong, Jiangsu Province 226001, China.
Biochim Biophys Acta Gen Subj. 2019 Mar;1863(3):598-608. doi: 10.1016/j.bbagen.2018.12.008. Epub 2018 Dec 17.
α1,6-Fucosyltransferase-deficient (Fut8) mice displayed increased locomotion and schizophrenia-like behaviors. Since neuroinflammation is a common pathological change in most brain diseases, this study was focused on investigating the effects of Fut8 in microglia and astrocytes.
Brain tissues were analyzed using immunohistochemical staining. Core fucosylation and protein expression were analyzed using lectin blot and western blot, respectively. Fut8-knockout (KO) cells were established by the CRISPR/Cas9 system.
The number of Iba-1 positive cells and GFAP positive cells were significantly increased in both untreated and lipopolysaccharide stimulated inflammatory conditional Fut8 mice by comparison with both wild-type (Fut8) and hetero (Fut8) mice. Stimulation with pro-inflammatory factors, such as IFN-γ and IL-6, induced expression levels of fucosylation in primary microglia and astrocytes, as well as in glial cell lines. Cell motility and iNOS expression were easily induced by IFN-γ in Fut8-KO BV-2 cells compared with wild-type (WT) cells. In a similar manner, both Fut8-KO C6 cells and primary astrocytes treated with 2-fluoro-L-fucose, a specific inhibitor for fucosylation, showed a higher response to IL-6-stimulated phospho-STAT3 signaling, compared with WT cells.
Core fucosylation negatively regulates the states of neuroinflammation by modulating the sensitivity of microglia and astrocytes to inflammatory mediators. The disorders of Fut8 mice are caused not only by neurons but also by glial cell dysfunction.
Core fucose is a novel regulator for neuroinflammation in the central nervous system.
α1,6-岩藻糖基转移酶缺陷（Fut8）小鼠表现出运动增加和类似精神分裂症的行为。由于神经炎症是大多数脑部疾病的共同病理变化，因此本研究集中于研究 Fut8 在小胶质细胞和星形胶质细胞中的作用。
使用免疫组织化学染色分析脑组织。使用凝集素印迹和 Western blot 分别分析核心岩藻糖基化和蛋白质表达。通过 CRISPR/Cas9 系统建立 Fut8 敲除（KO）细胞。
与野生型（Fut8）和杂合型（Fut8）小鼠相比，未经处理和脂多糖刺激的 Fut8 炎症条件小鼠的 Iba-1 阳性细胞和 GFAP 阳性细胞数量均显著增加。促炎因子（如 IFN-γ 和 IL-6）刺激诱导原代小胶质细胞和星形胶质细胞以及神经胶质细胞系中的岩藻糖基化表达水平。与野生型（WT）细胞相比，IFN-γ 容易诱导 Fut8-KO BV-2 细胞的细胞迁移和 iNOS 表达。类似地，与 WT 细胞相比，用 2-氟-L-岩藻糖（一种岩藻糖基化的特异性抑制剂）处理的 Fut8-KO C6 细胞和原代星形胶质细胞对 IL-6 刺激的磷酸化 STAT3 信号均表现出更高的反应。
核心岩藻糖基化通过调节小胶质细胞和星形胶质细胞对炎症介质的敏感性来负调控神经炎症状态。 Fut8 小鼠的紊乱不仅由神经元引起，也由神经胶质细胞功能障碍引起。
核心岩藻糖是中枢神经系统神经炎症的一种新的调节剂。
