镍离子诱导小鼠成纤维细胞氧化应激反应的实验研究

The study of nickel ions which induced oxidative stress in L929 cells

目的：研究镍离子对小鼠成纤维细胞L929的毒性作用规律及与氧化应激反应的关系。方法：小鼠成纤维细胞L929在不同镍离子浓度（0、200、400、800μmol／L）的培养液中培养24、48、72h，MTT法测定细胞相对增值率（relative growth rate，RGR）；培养48h，利用荧光探针DCFH-DA结合流式细胞术检测细胞内活性氧（reactive oxygen species，ROS）含量，通过比色法测定脂质过氧化反应的终产物丙二醛（maleic dialdehyde，MDA）。结果：200～800μmol／L镍离子处理细胞不同时间点下各组间存在统计学差异（P〈0．05）。各浓度组均表现出细胞毒性，中浓度组及高浓度组细胞毒性程度较高。200～800μmol／L镍离子处理细胞48h，各组间DCF荧光值及MDA生成量存在统计学差异（P〈0．05），且浓度越高，DCF荧光值及MDA生成量越高。结论：在镍离子刺激下，L929细胞增殖活力受到明显抑制，且呈现浓度依赖和时间依赖趋势；一定浓度的镍离子可诱导L929细胞发生氧化应激反应，氧化应激反应可能是镍离子细胞毒性反应的重要机制之一。

Objective： The object of this study was to investigate the cytotoxic effect of nickel ions and explore its relationship with ox- idative stress. Methods： L929 cells were exposed to different concentrations of nickel ions （0, 200,400, 800 μmol/L）for 24,48 and 72 h. MTT assay was used to assess the cytotoxicity of nickel ions. Cellular oxidative stress was evaluated by intracellular reactive oxygen species （ROS） production and the degree of lipid peroxidation （LPO） indicated by the levels of malondialdehyde （MDA）. Results： The results of MTT assay showed that cell viabilities at different culture time points were higher than those of the control. The intracellular ROS production and the content of MDA also increased in dose-dependent manner, which had significant difference to the control （P 〈 0.05 ）. Conclusions： From these studies, it was shown that cell proliferation of L929 cell was obviously inhibited by nickel ions. Nickel ions may induce intracellular oxidative stress, which may be one of mechanisms of cytotoxic effect of nickel ions on L929 cells.