目的探讨漆黄素(Fisetin,FIS)对小鼠巨噬细胞吞噬功能、NO的释放及对T淋巴细胞体外活化、增殖的影响。方法无菌制备小鼠巨噬细胞悬液及淋巴细胞悬液;荧光微球结合流式细胞术(FCM)分析FIS对巨噬细胞吞噬作用的影响;Griess试剂盒检测巨噬细胞NO的释放;双色荧光抗体染色结合FCM,检测CD3+T细胞CD69的表达水平;CFSE标记技术检测T细胞增殖的情况。结果 2.5μmol/L,5μmol/L,10μmol/LFIS均能明显抑制巨噬细胞的吞噬微球的能力;FIS能抑制LPS和IFN-γ刺激的巨噬细胞的NO的产生(P〈0.05);FIS对ConA刺激的T细胞表达CD69有抑制作用,并能有效抑制ConA诱导的T细胞增殖(P〈0.01),且均呈剂量依赖性。结论 FIS能显著抑制小鼠腹腔巨噬细胞的吞噬能力和分泌NO的能力,并能够抑制T细胞的活化和增殖,有望发展成为一种新的免疫抑制药物。
This study aims to investigate the effects of fisetin (FIS) on the mouse macrophages and T lymphocytes in vitro.Firstly,macrophages and lymphocytes of mice were prepared,and then stimulated with Fisetin.Fluorescence microbeads plus flow cytometry were used to analyze phagocytosis of macrophages of mouse;NO secretion of macrophages was detected by Griess reagent system;the expression of CD69 on CD3+T lymphocytes was measured by flow cytometry combined with two colored monoclonal antibodies;the effect of FIS on the proliferation of T lymphocytes was determined by CFSE staining technology.We found that NO production and phagocytosis rate of macrophages were inhibited by Fisetin (P 0.05);the expression of CD69 was significantly inhibited by Fisetin (P 0.01).Our results also showed that FIS (2.5 μmol/L,5 μmol/L and 10 μmol/L) significantly inhibited the proliferation of T lymphocytes(P 0.01).These data suggest that FIS can significantly inhibit the NO production and phagocytosis of macrophages,as well as can inhibit the proliferation and activation of T lymphocytes,thus promising to be developed as an immusuppressive drug.