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首页> 外文期刊>Neuroimmunomodulation >Lipopolysaccharide-Stimulated Transglutaminase 2 Expression Enhances Endocytosis Activity in the Mouse Microglial Cell Line BV-2
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Lipopolysaccharide-Stimulated Transglutaminase 2 Expression Enhances Endocytosis Activity in the Mouse Microglial Cell Line BV-2

机译:脂多糖刺激的转谷氨酰胺酶2表达增强小鼠小胶质细胞系BV-2的内吞作用。

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Objectives: In peripheral macrophages, tissue-type transglutaminase (TG2) is reported to be involved in phagocytosis of apoptotic cells. However, the contribution of TG2 to microglial phagocytosis has not been investigated. In this study, using a microglial cell line, BV-2, we examined the changes in TG2 expression, phagocytosis and pinocytosis in cells stimulated by lipopolysaccharide (LPS). Methods: Cells of the mouse microglial cell line BV-2 were stimulated by LPS with or without cystamine, an inhibitor of TG enzyme activity, for 24 h. TG2 expression was measured by real-time RTPCR and Western blotting. TG activity was evaluated using biotinylated pentylamine as a substrate. Pinocytosis was determined by uptake of 1-mu m fluorescent microbeads. Phagocytosis was assessed by uptake of dead cells, human neuroblastoma SH-SY5Y cells, which were pretreated with H2O2 for 24 h. Results: Phagocytosis of dead cells and pinocytosis of fluorescent microbeads were up-regulated by LPS stimulation together with TG2 expression. Blockade of TG enzyme activity by cystamine suppressed TG2 expression, phagocytosis and pinocytosis. Conclusions: These results suggested that LPS-induced TG2 was involved in the mechanism of pinocytosis and phagocytosis in microglia. (C) 2014 S. Karger AG, Basel
机译:目的:在外周巨噬细胞中,据报道组织型转谷氨酰胺酶(TG2)参与凋亡细胞的吞噬作用。但是,尚未研究TG2对小胶质细胞吞噬作用的贡献。在这项研究中,我们使用小胶质细胞系BV-2检查了脂多糖(LPS)刺激的细胞中TG2表达,吞噬作用和胞饮作用的变化。方法:LPS刺激小鼠小胶质细胞系BV-2的细胞,添加或不添加胱胺酸(一种TG酶活性的抑制剂)24小时。 TG2表达通过实时RTPCR和蛋白质印迹法测量。使用生物素化戊胺作为底物评估TG活性。通过摄取1μm的荧光微珠来确定胞饮作用。吞噬作用通过摄取死细胞,人类神经母细胞瘤SH-SY5Y细胞进行评估,死细胞用H2O2预处理24小时。结果:LPS刺激和TG2表达上调了死细胞吞噬作用和荧光微珠的胞饮作用。胱胺能阻断TG酶的活性,从而抑制TG2的表达,吞噬作用和胞饮作用。结论:这些结果表明LPS诱导的TG2参与了小胶质细胞的胞饮作用和吞噬作用。 (C)2014 S.Karger AG,巴塞尔

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