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目的应用双向电泳及串联质谱技术,研究不同剂量三氯乙烯(TCE)诱导 L-02肝细胞蛋白质表达谱的差异。方法 L-02肝细胞分别用不同剂量 TCE 和溶剂对照(DMSO)处理后,提取细胞总蛋白,双向凝胶电泳分离蛋白质组成分,银染显色,经 ImagerMaster 2D Platinum 5.0软件分组分析比较四组图谱,并对差异蛋白斑点进行基质辅助激光解吸/电离飞行时间串联质谱(MALDI-TOF-TOF-MS)分析鉴定。结果得出差异较明显的蛋白质斑点15个,不同剂量 TCE 刺激后表达分别上调、下调或消失,提示 TCE 可引起肝细胞蛋白表达发生改变。通过质谱鉴定和 IPI human 数据库检索,鉴定了其中9个 TCE 诱导 L-02肝细胞相关的蛋白质。结论这些结果为深入研究 TCE 毒作用相关的蛋白质及 TCE 毒作用的分子标志物提供依据,并为阐明 TCE 毒作用的分子机制提供线索。
Objective To study the difference of protein expression profiles of L-02 hepatocytes induced by different doses of trichlorethylene (TCE) using two-dimensional electrophoresis and tandem mass spectrometry. Methods The hepatocytes were treated with different doses of TCE and DMSO, respectively, and the total cellular protein was extracted. The proteome components were separated by two-dimensional gel electrophoresis and stained with silver stain. The four groups were analyzed by ImagerMaster 2D Platinum 5.0 software package At the same time, the spots of differentially expressed proteins were identified by MALDI-TOF-MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry). The results showed that there were 15 protein spots with obvious differences. The expression of TCE in different doses of TCE was up-regulated, down-regulated or disappeared respectively, suggesting that TCE could induce the change of hepatocyte protein expression. Through mass spectrometry identification and IPI human database search, nine TCE-induced L-02 hepatocyte-related proteins were identified. Conclusions These results provide the basis for further study on the TCE toxic effect-related proteins and molecular markers of TCE toxicity and provide clues for elucidating the molecular mechanism of TCE toxicity.