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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。
Objective To investigate the effect of Fisetin (FIS) on the phagocytosis of macrophages, the release of nitric oxide and the activation and proliferation of T lymphocytes in vitro. Methods The macrophage suspension and lymphocyte suspension were prepared aseptically. The effect of FIS on the phagocytosis of macrophages was analyzed by fluorescence microscopy combined with flow cytometry (FCM). The release of NO from macrophages was detected by Griess kit. The expression of CD69 on CD3 + T cells was detected by two-color fluorescent antibody staining combined with FCM. The proliferation of T cells was detected by CFSE labeling. Results FIS inhibited the phagocytosis of macrophages by 2.5μmol / L, 5μmol / L and 10μmol / L ascorbic acid. FIS inhibited the production of NO by macrophages stimulated by LPS and IFN-γ (P <0.05). FIS inhibited CD69 expression on ConA-stimulated T cells and inhibited ConA-induced T cell proliferation (P <0.01) in a dose-dependent manner. Conclusion FIS can significantly inhibit the phagocytosis and secretion of NO peritoneal macrophages in mice, and can inhibit the activation and proliferation of T cells. It is expected to develop into a new immunosuppressive drug.