缺氧对巨噬细胞HMGB1释放的影响及机制

发布时间：2018-06-26 01:35

本文选题：缺氧 + 高迁移率族蛋白B1　；参考：《南京医科大学》2012年硕士论文

【摘要】：目的观察缺氧对巨噬细胞高迁移率族蛋白B1（HMGB1）释放的诱导作用，，并探讨其可能机制。方法采用巨噬细胞株RAW264.7，观察缺氧（1%O2或100μmol/LCoCl2）培养不同时间（6、12、24h）对细胞培养上清液中HMGB1含量、细胞HMGB1mRNA及细胞HMGB1胞内分布的影响。观察不同浓度N-乙酰半胱氨酸（NAC）、MAPK信号通路抑制剂(PD98059、SP600125、SB202190)对缺氧诱导细胞HMGB1释放的影响。HMGB1上清液含量和HMGB1mRNA表达水平分别采用酶联免疫吸附试验（ELISA）、实时荧光定量PCR（RT-PCR）检测。结果巨噬细胞株RAW264.7缺氧（1%O2或100μmol/LCoCl2）培养12h后培养上清液中HMGB1含量及细胞HMGB1mRNA表达升高（P0.05或P0.01），24h时达到峰值；缺氧（1%O2）培养12h时，免疫荧光染色显示HMGB1出现核浆转移；NAC（10、100mmol/L）、PD98059（5、20μmol/L）、SP600125（5、20μmol/L）对巨噬细胞株RAW264.7缺氧（1%O2）培养24h后培养上清液中的HMGB1含量有明显抑制作用（P0.01）。结论缺氧能诱导巨噬细胞释放HMGB1，其机制可能与胞内氧自由基产生及MEKJNK信号通路有关。
[Abstract]:Objective to investigate the effect of hypoxia on the release of high mobility group protein B1 (HMGB1) from macrophages and its possible mechanism. Methods macrophage cell line RAW264.7 was used to observe the effects of hypoxia (1O 2 or 100 渭 mol / L CoCl2) on HMGB1 content, HMGB1 mRNA and intracellular distribution of HMGB1 in supernatant of cell culture. The effects of different concentrations of N-acetylcysteine (NAC) MAPK signaling pathway inhibitor (PD98059 / SP600125 / SB20190) on hypoxia induced HMGB1 release were investigated by enzyme linked immunosorbent assay (Elisa) and real-time fluorescence quantitative PCR (RT-PCR). Results the HMGB1 content and HMGB1 mRNA expression in the supernatant of macrophages RAW264.7 exposed to hypoxia (1O 2 or 100 渭 mol / L CoCl2) reached a peak at 24 h after incubation (P0.05 or P0.01), and the nuclear and cytoplasmic metastasis of HMGB1 was detected by immunofluorescence staining at 12 h after hypoxia (1O2). NAC (10 100 mmol / L) PD98059 (5 ~ 20 渭 mol / L) SP600125 (5 ~ 20 渭 mol / L) inhibited the HMGB1 content in the supernatant of RAW264.7 cultured for 24 h after hypoxia (1O2) (P0.01). Conclusion hypoxia can induce the release of HMGB1 from macrophages, which may be related to the production of intracellular oxygen free radicals and MEKJNK signaling pathway.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.11

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