目的 研究螺环内酯型对映-贝壳杉烷型二萜rabdosin B （RB）诱导HL-60向成熟粒细胞分化的特征及机制。方法 采用台盼蓝染色、吉姆萨染色、硝基四氮唑蓝（NBT）还原力测定及流式细胞术检测RB对HL-60细胞增殖、细胞周期、NBT还原力、细胞吞噬及细胞表面抗原CD11b表达的影响。结果 RB在1.0、3.0、5.0 μmol/L下抑制HL-60细胞增殖，引起HL-60细胞G₀/G₁期阻滞，增加HL-60细胞肾形核和分叶核细胞比率，并伴随细胞NBT还原力和吞噬能力显著增强以及CD11b阳性细胞率明显上升。进一步检测显示，活性氧（ROS）清除剂N-乙酰-L-半胱氨酸（NAC，300 μmol/L）及NADPH氧化酶抑制剂夹竹桃麻素（APO，100 μmol/L）和二联苯碘锚（DPI，0.1 μmol/L）不仅可显著抑制RB诱导的细胞内ROS升高，并且可显著抑制RB诱导的HL-60细胞分化的各项特征指标的改变。结论 RB可通过上调HL-60细胞NADPH氧化酶活性，升高胞内ROS浓度，诱导HL-60细胞向成熟粒细胞分化。

Objective To investigate the characteristics and mechanism of spironolactone ent-kaurane diterpene type diterpene rabdosin B (RB) induced HL-60 differentiation into mature granulocytes.Methods The effects of RB on proliferation, cell cycle, NBT reducing power, phagocytosis, and cell surface antigen CD11b expression of HL-60 were detected by trypan blue staining, Giemsa staining, NBT reducing power assay, and flow cytometry, respectively.Results RB inhibited the proliferation of HL-60 cells at the concentration of 1.0, 3.0, and 5.0 μmol/L, which resulted in the arrest of G₀/G₁ phase, increased the ratio of renal nucleated and lobate nucleus, and markedly enhanced the NBT reducing power and phagocytic capacity and the significant increase of CD11b positive cell rate. Further tests showed that the active oxygen scavenger NAC (300 μmol/L), NADPH oxidase inhibitor APO (100 μmol/L), and DPI (0.1 μmol/L) not only significantly inhibited the intracellular ROS upregulation induced by RB, but also inhibited various features of RB induced HL-60 cell differentiation.Conclusion RB can increase the activity of NADPH oxidase in HL-60 cells, increase the intracellular ROS concentration, and induce the differentiation of HL-60 cells to mature granulocytes.

国家自然科学基金资助项目（31660101，30960464）