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目的:以H_2O_2造成人白血病HL-60细胞的氧化损伤,研究含硒化合物Ebselen(Ebs)对氧化性损伤的影响。方法:MTT比色法测定细胞增殖,TBA法检测膜脂过氧化水平,单细胞电泳法确定DNA损伤程度,并用荧光探针DCFH-DA检测细胞内活性氧自由基(ROS)水平的变化。结果:H_2O_2(100μmol·L~(-1)~)可显著抑制HL-60细胞增殖,引起膜脂过氧化水平升高,Ebs对此表现出浓度依赖性的抑制效应,Ebs20μmol·L~(-1)对H_2O_2 100μmol·L~(-1)引起的膜脂过氧化水平抑制率达56.4％;同样,Ebs对H_2O_2 100μmol·L~(-1)造成的DNA损伤和细胞内活性氧水平的升高均有拮抗效应,也呈明显量效关系。结论:含硒化合物Ebs对由活性氧诱发的细胞毒性和DNA损伤有强的防护效应。
OBJECTIVE: To study the oxidative damage of human leukemia HL-60 cells induced by H 2 O 2 and the effect of selenium-containing compound Ebselen (Ebs) on oxidative damage. Methods MTT assay was used to measure the cell proliferation. The level of membrane lipid peroxidation was determined by TBA method. The degree of DNA damage was determined by single cell electrophoresis. The level of reactive oxygen species (ROS) in cells was detected by fluorescent probe DCFH-DA. Results: H 2 O 2 (100μmol·L -1) ~ (-1) could significantly inhibit the proliferation of HL-60 cells and cause the increase of membrane lipid peroxidation. Ebs showed a concentration-dependent inhibitory effect. Ebs 20μmol·L ~ 1) inhibited the membrane lipid peroxidation induced by H 2 O 2 100μmol·L -1 up to 56.4%. Similarly, the effect of Ebs on DNA damage and intracellular reactive oxygen species rise induced by H 2 O 2 100μmol·L -1 High antagonistic effect, but also showed a significant dose-effect relationship. CONCLUSIONS: The selenium compound Ebs has a strong protective effect against cytotoxicity and DNA damage induced by reactive oxygen species.