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目的研究金雀异黄素对巨噬细胞RAW264.7增殖、细胞形态及细胞周期的影响,以探讨其对免疫系统的作用。方法以鼠源巨噬细胞系RAW264.7为细胞模型,将金雀异黄素用二甲基亚砜(DMSO)溶解,再用培养液稀释成所需终浓度为12.5、25、50、100μmol·L-1的4组及溶剂对照组(DMSO体积含量比低于0.1%)。通过WST-1细胞增殖实验、倒置相差显微镜及流式细胞仪分别检测金雀异黄素对巨噬细胞的增殖、细胞形态及细胞周期的影响。结果金雀异黄素对巨噬细胞的影响呈时间和剂量依赖关系。50～100μmol·L-1浓度的金雀异黄素作用巨噬细胞24h和48h后均能抑制细胞增殖,细胞增殖率分别为77%～81%、58%～73%(P<0.01);并能影响细胞形态改变,表现为细胞体积增大,并形成较长伪足。流式细胞结果显示,该浓度金雀异黄素可将巨噬细胞阻滞在S-G2/M期。结论金雀异黄素能抑制巨噬细胞细胞增殖,影响巨噬细胞形态,可能与干扰细胞周期有关。
Objective To investigate the effects of genistein on the proliferation, cell morphology and cell cycle of RAW264.7 macrophages in order to investigate its effect on the immune system. Methods Rat macrophage cell line RAW264.7 was used as a cell model. Genistein was dissolved in dimethylsulfoxide (DMSO) and diluted to the desired final concentration of 12.5, 25, 50 and 100 μmol 4 groups of L-1 and solvent control group (DMSO volume content less than 0.1%). The effects of genistein on proliferation, cell morphology and cell cycle of the macrophages were detected by phase contrast microscopy and flow cytometry, respectively, through WST-1 cell proliferation assay. Results genistein on macrophages in a time-and dose-dependent manner. The genistein treated with genistein at a concentration of 50 ~ 100μmol·L-1 all inhibited cell proliferation 24h and 48h, with the cell proliferation rates of 77% -81% and 58% -73%, respectively (P <0.01). And can affect cell morphology changes, manifested as cell volume increases, and the formation of longer pseudo-foot. Flow cytometry results showed that this concentration of genistein blocked the macrophages in the S-G2 / M phase. Conclusion Genistein can inhibit the proliferation of macrophages and affect the morphology of macrophages, which may be related to the interference of cell cycle.