高级检索
    苏静, 孔庆玲, 李淼, 祁继金, 仲青, 周竟心, 高玲, 胡娜, 陈伟. LPS刺激RAW264.7、THP-1细胞HMGB1释放及共刺激分子表达[J]. 徐州医科大学学报, 2022, 42(11): 799-804. DOI: 10.3969/j.issn.2096-3882.2022.11.004
    引用本文: 苏静, 孔庆玲, 李淼, 祁继金, 仲青, 周竟心, 高玲, 胡娜, 陈伟. LPS刺激RAW264.7、THP-1细胞HMGB1释放及共刺激分子表达[J]. 徐州医科大学学报, 2022, 42(11): 799-804. DOI: 10.3969/j.issn.2096-3882.2022.11.004

    LPS刺激RAW264.7、THP-1细胞HMGB1释放及共刺激分子表达

    • 摘要: 目的 探讨细菌脂多糖(LPS)对巨噬细胞高迁移率族蛋白B1(HMGB1)及共刺激分子表达的影响。方法 培养THP-1、RAW264.7细胞,予以不同浓度的LPS刺激24h,CCK8法检测细胞增殖的变化;流式细胞术检测细胞凋亡;免疫印迹法检测培养上清以及细胞内HMGB1的水平;定量聚合酶链反应(Q-PCR)检测细胞内HMGB1转录水平;流式细胞术检测细胞表面CD80、CD83及CD86的表达的变化。结果 LPS 促进RAW264.7增殖,低浓度LPS抑制THP-1增殖,高浓度LPS促进THP-1细胞增殖;LPS促进THP-1、RAW264.7两种细胞的凋亡;THP-1及RAW264.7细胞的培养上清中的HMGB1均增加(P<0.05),细胞内HMGB1水平下降(P<0.05)。THP-1、RAW264.7细胞内HMGB1的mRNA转录水平升高(P<0.05);流式细胞术检测THP-1、RAW264.7细胞表面CD80、CD83、CD86的表达示LPS促进细胞表面CD80、CD83和CD86的表达(P<0.05)。结论 LPS 影响巨噬细胞增殖和凋亡,引起细胞内HMGB1分泌至胞外,促进细胞表面共刺激分子表达增加。

       

      Abstract: ob<x>jective To investigate the effect of bacterial lipopolysaccharide (LPS) on the ex<x>pression of macrophage high mobility group protein B1(HMGB1) and costimulatory molecules. Methods THP-1 and RAW264.7 cells were cultured and stimulated with LPS of different concentrations for 24h. The changes of cell proliferation were detected by CCK8 method . Flow cytometry were used to test the level of apoptosis.The Level of HMGB1 in the culture supernatant and cell was detected by western blot. Q-PCR tested the level of HMGB1 transc<x>ription. The ex<x>pression of CD80, CD83, and CD86 were detected by flow cytometry. Results CCK8 assay showed that LPS promoted the proliferation of RAW264.7. By contrast, low concentration LPS restrained THP-1 proliferation, while high concentration LPS promoted THP-1 proliferation (P<0.05). Flow cytometry detected that LPS promoted cell apoptosis (P<0.05). Western blot showed the ex<x>pression of HMGB1 in THP-1 and RAW264.7 cells culture supernatant raised (P<0.05), while the intracellular HMGB1 decreased (P<0.05). The transc<x>riptional level of HMGB1 in THP-1 and RAW264.7 cells went up tested by Q-PCR (P<0.05). The ex<x>pression of CD80, CD83, and CD86 increased on these two cells(P<0.05). Conclusion LPS affects THP-1 and RAW264.7 cells proliferation and apoptosis, promotes HGMB1 release, increases the cell surface ex<x>pression of CD80, C883, and CD86.

       

    /

    返回文章
    返回