甲醛诱导mrc5细胞适应性反应蛋白质研究.docVIP

甲醛诱导MRC5细胞适应性反应蛋白质研究 作者：李习艺，庄志雄，刘建军，杨晓华，卫秦芝，黄海燕 【关键词】 甲醛；适应性反应；蛋白质组 　　摘要： 目的 研究甲醛诱导真核细胞适应性反应及其可能的机制。方法 建立甲醛刺激人胚肺成纤维细胞(MRC-5)适应性反应模型；采用AlamarBlue还原率检测细胞增殖。双向凝胶电泳分离细胞总蛋白，图像分析后选取差异蛋白点进行质谱鉴定。结果 AlamarBlue检测结果表明，低剂量甲醛预刺激可诱导细胞适应性反应。双向凝胶电泳结果显示，甲醛刺激后19个蛋白斑点发生变化，肽指纹图谱及肽序列标签鉴定了其中13个蛋白斑点，已鉴定的蛋白包括二磷酸核苷酸激酶、延长因子1-γ，磷酸丙糖异构酶、60S酸性核糖体蛋白P0、75kDa热休克蛋白、70kD热休克蛋白样结合蛋白、钙依赖磷脂结合蛋白I、假想蛋白FLJ34423、微管-肌动蛋白交叉连接因子1、核纤层蛋白B2、ATP合成酶α链、蛋白酶体α亚基6，这些蛋白功能涉及转录调节、蛋白折叠、信号传导、能量代谢、细胞骨架等各个方面。结论 低剂量甲醛可诱导MRC-5细胞适应性反应，多种蛋白参与细胞的适应性反应调控。 　　关键词： 甲醛；适应性反应；蛋白质组 　　Proteomie analysis of adaptive response induced by low concentration of formaldehyde in human embryonic fibroblasts cells 　　Abstract： Objective To study the adaptive response induced by formaldehyde(FA) on eukaryocyte and its possible mechanism.Methods After FA treatment，AlamarBlue reducing rate was observed to determine the cell proliferation.The total cellular proteins were separated using two-dimensional gel electrophoresis and visualized by sliver staining.Gels from different batches of control and FA-treated MRC-5 cells were analyzed for quantitative spot comparisons with the ImageMaster 2D platinum 5.0 software.The differentially expressed prorein spots were picked and digested in gel then identified by tandem mass spectrum.Results The results of AlamarBlue reducing rate showed that low concentration of FA induced adaptive response on MRC-5 cells.Nineteen spots were changed significantly after FA treatment.Thirteen proteins were identified by peptide mass fingerprinting or peptide sequence analysis，including putative nucleoside diphosphate kinase，elongation factor 1-gamma，triosephosphate isomerase，60S acidic ribosomal protein P0，heat shock protein 75 kDa,similar to heat shock 70kD protein binding protein，annexin I，hypothetical protein FLJ34423，microtubule-actin crosslinking factor 1，lamin B2，ATP synthase alpha chain，mitochondrial precursor，proteasome subunit alpha type 6.These identified proteins involved