O-linked N-acetylglucosamine glycosylation of p65 aggravated the inflammation in both fibroblast-like synoviocytes stimulated by tumor necrosis factor-α and mice with collagen induced arthritis.
O-linked N-acetylglucosamine glycosylation of p65 aggravated the inflammation in both fibroblast-like synoviocytes stimulated by tumor necrosis factor-α and mice with collagen induced arthritis.
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p65的O连接N-乙酰葡萄糖糖基化加剧了肿瘤坏死因子-α刺激的成纤维细胞样的滑膜细胞和胶原蛋白诱导的关节炎刺激的炎症。
DOI：
10.1186/s13075-015-0762-7
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发表时间：
2015-09-14
影响因子：
4.9
通讯作者：
Cho JW
中科院分区：
文献类型：
--
作者：
Kim HB;Lee SW;Mun CH;Yoon JY;Pai J;Shin I;Park YB;Lee SK;Cho JW
我们研究了类风湿性关节炎（RA）中p65的O-连接N-乙酰葡萄糖胺糖基化（O-GlcNAc化）的炎症潜力。用合成ThiaMet-G（200 μM）（一种O-GlcNAcase（OGA）抑制剂）处理成纤维细胞样滑膜细胞（FLS）和MH 7A细胞，然后用肿瘤坏死因子（TNF）-α（10 μg/mL）处理。采用四甲基偶氮唑盐（MTT）法检测滑膜细胞的增殖情况，采用RT-PCR法定量检测编码促炎分子的mRNA水平。对p65的O-GlcNAc化的核定位、DNA结合亲和力和转录活性进行了评估。对胶原诱导的关节炎（CIA）小鼠进行关节炎严重程度评估和组织病理学检查。ThiaMet-G（20 mg/kg）腹腔注射，隔日1次，共26 d。进行T细胞的活化细胞分选（FACS）分析。高-O-GlcNAc酰化增加TNF-α刺激的滑膜细胞增殖和促炎基因mRNA表达。此外，O-GlcNAc修饰增强了p65的核定位比例、DNA结合亲和力和转录活性。在CIA小鼠中，ThiaMet-G在临床和组织学上显著加重关节炎的严重程度，并且还增加了CD 4 + IFN-γ + T细胞和CD 4 + IL-17+ T细胞。p65的O-GlcNAc化增加了TNF-α介导的炎症在体外（滑膜细胞）和体内（CIA小鼠）的作用。本文的在线版本（doi：10.1186/s13075-015-0762-7）包含补充材料，可供授权用户使用。
We investigated the inflammatory potential of O-linked N-acetylglucosamine glycosylation (O-GlcNAcylation) of p65 in rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) and MH7A cells were treated with synthetic ThiaMet-G (200 μM), an O-GlcNAcase (OGA) inhibitor, followed by tumor necrosis factor (TNF)-α (10 μg/mL). Proliferation of synovial cells was measured by MTT assay, and the levels of mRNAs encoding pro-inflammatory molecules were quantitated by RT-PCR. The nuclear localization of O-GlcNAcylated of p65 and its DNA binding affinity and transcriptional activity were assessed. The severity assessment of arthritis and a histopathological examination were done in mice with collagen induced arthritis (CIA). ThiaMet-G (20 mg/kg) intraperitoneal injection was done every other day for 26 days. Fluorescence-activated cell sorting (FACS) analysis of T cells was performed. Hyper-O-GlcNAcylation increased the proliferation and mRNA expression of pro-inflammatory genes in synoviocytes stimulated by TNF-α. Moreover, O-GlcNAcylation of p65 enhanced its proportion of nuclear localization, DNA binding affinity and transcriptional activity. In CIA mice, ThiaMet-G significantly aggravated the severity of arthritis clinically and histologically, and it also increased CD4 + IFN-γ + T cells and CD4 + IL-17+ T cells. O-GlcNAcylation of p65 increased the effects of TNF-α-mediated inflammation both in vitro (in synovial cells) and in vivo (in mice with CIA). The online version of this article (doi:10.1186/s13075-015-0762-7) contains supplementary material, which is available to authorized users.