Effects of mimicked acetylated HMGB1 on macrophages and dendritic cells.
Effects of mimicked acetylated HMGB1 on macrophages and dendritic cells.
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模拟乙酰化 HMGB1 对巨噬细胞和树突状细胞的影响
DOI：
10.3892/mmr.2018.9584
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发表时间：
2018-12
影响因子：
3.4
通讯作者：
Zheng F
中科院分区：
文献类型：
--
作者：
Chen X;Xu Y;Xiong P;Tan Z;Gong F;Hou X;Zheng F
细胞外高迁移率族蛋白1（HMGB 1）在炎症性疾病中起关键作用。HMGB 1主要通过坏死细胞的被动释放或刺激后单核细胞/巨噬细胞的主动分泌释放到细胞外环境中。然而，需要通过翻译后修饰（如乙酰化）将活跃分泌的HMGB 1从细胞核转运至细胞质;然后将HMGB 1释放至细胞外间隙。乙酰化是否影响HMGB 1的细胞外功能仍不清楚。本研究通过设计引物，优化了基因突变的方法，并对突变基因进行了鉴定。用6个赖氨酸取代谷氨酰胺模拟乙酰化HMGB 1（HMGB 1-M），观察HMGB 1-M对巨噬细胞和树突状细胞（DCs）的作用。用ELISA法检测RAW 264.7细胞中肿瘤坏死因子（TNF）-α的产生。流式细胞术检测RAW 264.7细胞吞噬能力、DC成熟和CXCR 4表达。本研究的结果表明，HMGB 1-M增加胞质易位。与HMGB 1相比，HMGB 1-M可增加RAW 264.7细胞内TNF-α的产生，降低DCs上整合素α X的平均荧光强度（MFI）和主要组织相容性复合物-II的百分比及MFI。HMGB 1-M对巨噬细胞的吞噬功能及DC上分化抗原80和趋化因子受体4的表达频率无明显影响。提示HMGB 1-M可部分促进炎症反应，降低DC成熟。因此，本研究的结果可能会提供深入了解HMGB 1在炎症性疾病中的复杂作用。
Extracellular high mobility group box 1 (HMGB1) serves a critical role in inflammatory diseases. HMGB1 is released into the extracellular environment mainly by passive release from necrotic cells or active secretion from monocytes/macrophages following stimulation. However, the translocation of actively secreted HMGB1 from the nucleus to the cytoplasm with post-translational modifications such as acetylation is required; HMGB1 is then released into the extracellular space. Whether acetylation influences the extracellular function of HMGB1 remains unknown. In the present study, an optimized method of gene mutation by using well-designed primers in particular, which were employed to identify the mutant gene. The substitution of six lysine residues for glutamines was conducted to mimic acetylated HMGB1 (HMGB1-M) and observe the effects of HMGB1-M on macrophages and dendritic cells (DCs). Tumor necrosis factor (TNF)-α production in RAW 264.7 cells was assessed by ELISA. The phagocytic potential of RAW 264.7 cells, DC maturation and CXCR4 expression were analyzed by flow cytometry. The results of the present study revealed that HMGB1-M increased cytoplasmic translocation. Compared with HMGB1, HMGB1-M increased TNF-α production within RAW 264.7 cells and decreased the mean fluorescence intensity (MFI) of integrin α X, and the percentage and MFI of major histocompatibility complex-II on DCs. HMGB1-M exhibited no significant effects on phagocytosis of macrophages and expression frequency of cluster of differentiation 80 and chemokine receptor type 4 on DCs. These results suggested that HMGB1-M may partly promote inflammation and decrease DC maturation. Thus, the findings of the present study may provide insight into the complex role of HMGB1 in inflammatory diseases.
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影响因子：
64.8
作者：
Scaffidi, P;Misteli, T;Bianchi, ME
通讯作者：
Bianchi, ME
影响因子：
2.2
作者：
Lutz, MB;Kukutsch, N;Schuler, G
通讯作者：
Schuler, G
影响因子：
4.1
作者：
Assenberg, Ren;WebB, Michelle;Thomas, Jean O.
通讯作者：
Thomas, Jean O.
DOI：
10.1007/978-1-62703-293-3_13
发表时间：
2013-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Hsieh, Pei-Chung;Vaisvila, Romualdas
通讯作者：
Vaisvila, Romualdas
DOI：
10.1016/j.bbagrm.2009.11.009
发表时间：
2010-01
影响因子：
4.7
作者：
Zhang, Qingchun;Wang, Yinsheng
通讯作者：
Wang, Yinsheng