摘要目的 探讨Embelin是否通过氧化应激途径导致DNA双链损伤,从而抑制HL-60细胞增殖.方法 HL-60细胞经不同浓度(3、10、30、100及300 μg/ml)Embelin处理24 h,采用CCK-8法检测细胞增殖抑制率;采用DCFH-DA荧光探针流式细胞术测定细胞内活性氧(ROS)水平;彗星试验检测DNA双链断裂(DSB)情况.结果 与对照组相比,Embelin浓度为10、30、100及300 μg/ml时均可显著抑制HL-60细胞增殖,增殖抑制率分别为(12.74±2.27)％、(23.49±1.96)％、(30.30±1.89)％和(57.55±3.59)％(P值均＜0.05);随着药物浓度的增加HL-60细胞内ROS的水平增高、DSB增加(p值均＜0.05);应用ROS抑制剂N-乙酰-L-半胱氨酸(NAC)预先处理HL-60细胞2h后再加入300 μg/ml的Embelin作用24 h,与单用300 μg/ml Embelin作用组比较,ROS水平下降、DSB减少,同时Embelin对HL-60细胞的抑制率由(57.55±3.59)％下降为(32.75±2.70)％(P值均＜0.05).结论 Embelin通过产生细胞内ROS发生氧化应激作用,导致DSB,最终抑制HL-60细胞的增殖.

abstractsObjective To evaluate the effects of Embelin on HL-60 cells by the impact of oxidative stress on DNA double-strain breaks(DSBs).Methods HL-60 cells were treated with Embelin in different concentration(3,10,30,100,and 300 tg/ml) for 24 h,and inhibitory effects was examined by CCK-8 assay.Reactive oxygen species (ROS) levels were evaluated by flow cytometry using DCFH-DA.Comet assay was used to detect the extent of DSBs.Results Embelin inhibited proliferation of HL-60 cells in a dose-dependent manner.At the concentration of 10,30,100,and 300 μg/ml,the inhibition rate was (12.74±2.27)％,(23.49±1.96)％,(30.30±1.89)％,and (57.55± 3.59)％ (P＜0.05).Embelin also lead to high level of intracellular ROS and deterioration of DNA damage (P＜0.05).When HL-60 cells were pretreated with ROS scavenger N-acetyl-l-cysteine(NAC) for 2 h and then treated with 300 μg/ml Embelin for 24 h,the intracellular ROS level declined and DSBs relieved (P＜0.05).Meanwhile,embelin-induced cell viability significantly declined to(32.75±2.70)％(P＜0.05).Conclusions Embelin induced the death of HL-60 cells by increasing the generation of intracellular oxidation and the oxidative stress,which drived the damage of DNA double-strand.