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探讨根皮素(phloretin)在体外对小鼠T淋巴细胞增殖、活化、周期和巨噬细胞NO释放、吞噬功能的影响。CFDA-SE标记技术结合流式细胞术检测根皮素对T淋巴细胞增殖的影响。荧光抗体染色结合流式细胞术分析根皮素对T淋巴细胞在ConA的刺激下CD69和CD25的表达水平的影响。碘化丙啶(PI)染色检测细胞周期的分布情况。Griess试剂盒和荧光微球分别用以检测巨噬细胞NO释放和吞噬功能。结果显示,根皮素(40、60和80μmol.L-1)明显抑制T淋巴细胞增殖,增殖指数(PI)从1.41±0.13分别下降到1.34±0.16、1.19±0.12和1.07±0.06。根皮素可明显抑制CD69和CD25的表达(P<0.01)。细胞周期分析显示,根皮素可将细胞抑制在G0/G1期。巨噬细胞在LPS和IFN-γ的刺激下的NO释放量为(26.72±3.57)μmol.L-1,而在根皮素作用下NO释放量明显下降(P<0.01)。巨噬细胞吞噬率在根皮素作用下也明显下降(P<0.01)。以上结果表明,根皮素有望发展成为一种新的免疫抑制药物。
To investigate the effects of phloretin on the proliferation, activation, cycle and macrophage NO release and phagocytosis of mouse T lymphocytes in vitro. The effect of phloretin on the proliferation of T lymphocytes was detected by CFDA-SE and flow cytometry. Fluorescent antibody staining combined with flow cytometry analysis of the effects of phloretin on the expression of CD69 and CD25 on T lymphocytes stimulated by ConA. Propidium iodide (PI) staining to detect cell cycle distribution. Griess kit and fluorescent microspheres were used to detect macrophage NO release and phagocytosis. The results showed that phloretin (40, 60 and 80 μmol·L-1) significantly inhibited the proliferation of T lymphocytes. The proliferation index (PI) decreased from 1.41 ± 0.13 to 1.34 ± 0.16, 1.19 ± 0.12 and 1.07 ± 0.06, respectively. Phloretin significantly inhibited the expression of CD69 and CD25 (P <0.01). Cell cycle analysis showed that phloretin inhibited cells in the G0 / G1 phase. The release of NO by macrophages stimulated by LPS and IFN-γ was (26.72 ± 3.57) μmol.L-1, while the NO release was significantly decreased by phloretin (P <0.01). Phagocytic rate of macrophages also decreased significantly under the action of phloretin (P <0.01). The above results indicate that phloretin is expected to develop into a new immunosuppressive drug.