姜黄素对人正常肝细胞角蛋白1和内质网蛋白19表达的影响

目的应用双向凝胶电泳、质谱技术和蛋白免疫印迹研究姜黄素(Curcumin,CUR)对人正常肝细胞蛋白质组的影响。方法随机将人正常肝细胞L-02分成两组,对照组不作任何处理,姜黄素组以25μmol.L-1姜黄素处理L-02细胞24 h。用双向凝胶电泳建立姜黄素组和对照组的双向凝胶电泳图谱,并用Image Master 2D Platinum 6.0软件进行差异蛋白质组分析,所获得的明显差异蛋白质用MALDI-TOF-MS进行鉴定分析,最后用蛋白免疫印迹法进行验证。结果姜黄素作用于人正常肝细胞后,有2个蛋白质的表达发生明显变化,角蛋白1与内质网蛋白19表达均上调。结论上调角蛋白1和内质网蛋白19可能是姜黄素抗肿瘤、抗氧化作用的机制之一。

Effects of curcumin on expression of cytokeratin 1 and endoplasmic reticulum protein 19 in human normal hepatocytes

Aim To observe the effects of curcumin(CUR) on human normal hepatocytes proteome by bidimensional gel electrophoresis,mass spectrometry and western blot.Methods The L-02 cells were randomly divided into two groups,control group and CUR group in which the cells were treated with CUR(25 μmol·L-1)for 24 hours.The bidirectional gel electrophoresis was used to establish the two-dimensional gel electrophoresis patterns of proteome in group of human normal hepatic cells exposed to CUR and in control group.Image Master 2D Platinum 6.0 software was used to analyse the differentially expressed proteins in two groups,and the matrix assisted laser desorption/ionization-time of flight-mass spectrometry(MALDI-TOF-MS) was used to identify them.Western blot was used to verify them.Results Among the differentially expressed proteins in the two groups,CK1 and ERp19 were up-regulated markedly;two proteins had been identified by MALDI-TOF-MS and western blot.Conclusion Up-regulation of CK1 and ERp19 may be one mechanism of antioxidant and anti-tumor effects of CUR.