NADPH氧化酶源性活性氧介导epinodosin诱导HL-60细胞分化及细胞骨架重组

NADPH Oxidase-derived ROS Mediated the Differentiation and Reorganization of Cytoskeletalon Induced by Epinodosin in HL-60 Cells

该文分析研究了对映-贝壳杉烷二萜epinodosin通过调节NADPH氧化酶源活性氧诱导急性早幼粒细胞白血病细胞HL-60向成熟粒细胞分化的特征。结果显示,epinodosin在4.0~8.0μmol/L浓度时抑制HL-60细胞生长,导致其S期阻滞,诱导HL-60细胞形成肾形核和多叶核细胞,使细胞吞噬能力和对硝基四氮唑蓝（nitrotetrazolium blue chloride,NBT）的还原力显著增强、细胞表面抗原CD11b表达上调,表明epinodosin可诱导HL-60细胞向成熟粒细胞分化;epinodosin还诱导HL-60细胞骨架重组,显著促进微管的成束组装以及提高波形纤维含量,上述分化特征与临床诱导分化治疗剂全反式维甲酸（all-trans retinoic acid,ATRA）对HL-60细胞的分化诱导特性极为相似。进一步检测显示,抗氧化剂NAC（N-acetyl-L-cysteine）以及NADPH氧化酶抑制剂夹竹桃麻素（apocynin,APO）和联苯基三价碘（diphenyleneiodonium,DPI）均可抑制epinodosin对HL-60细胞的分化诱导作用,表明epinodosin导致HL-60细胞活性氧浓度的升高与NADPH氧化酶活性关联,提示epinodosin通过调节NADPH氧化酶源性活性氧诱导HL-60向粒细胞分化。

In this paper, NADPH oxidase-derived reactive oxygen species（ROS） mediated the differentiation of HL-60 to mature granulocyte caused by epinodosin were studied. The results showed that the growth of HL-60 was inhibited by epinodosin under the concentration of 4.0~8.0 μmol/L, and it was arrested at S phase. Epinodosin induced HL-60 formed kidney-shaped nuclei and lobulated nuclei, enhanced nitrotetrazolium blue chloride（NBT）-reducing ability, cellular uptake and the expression of CD11 b of HL-60 cells. These indicated that epinodosin could induce the differentiation of human HL-60 cells to granulocyte. And also, epinodosin could induce the reorganization of cytoskeleton in HL-60 cells, improve the assembly of microtubules and vimentin filaments. All the differentiation characteristics mentioned above were similar as the results of clinical differentiation therapy all-trans retinoic acid（ATRA） acted on HL-60 cells. Further detection displayed that the inducement of differentiation by epinodosin was inhibited by N-acetyl-L-cysteine（NAC, an antioxidant） andapocynin/diphenyleneiodonium（APO/DPI, the inhibitors of NADPH oxidase）. Namely, the increase of ROS was associated with NADPH oxidase, it revealed NADPH oxidase-derived reactive oxygen species mediated HL-60 cells differentiation caused by epinodosin.