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目的探讨蛋白质组学技术研究二氧化硅粉尘处理的肺泡巨噬细胞(AM)上清液对人胚肺成纤维细胞(MRC-5)蛋白质表达变化的影响。方法实验组采用二氧化硅处理大鼠AM的培养上清液刺激MRC-5,对照组MRC-5采用等量无粉尘处理的AM培养上清液作用;提取MRC-5总蛋白,进行双向凝胶电泳识别差异表达蛋白,经质谱分析、数据库搜索进行蛋白鉴定。结果对照组MRC-5细胞检测到(410±21)个,染尘组为(398±19)个蛋白点。组间的匹配性较好,染尘上清刺激后35个蛋白斑点发生变化,初步鉴定出数个与氧化应激相关的蛋白:热休克蛋白(HSP27)、Per-oxiredoxinⅡ亚型b、钙网蛋白-3和钙网蛋白-5类似物等。结论应用蛋白质组学方法有助于筛选到经不同处理细胞间的蛋白表达差异,为进一步研究矽肺机制提供分子基础。
Objective To investigate the effects of silica dust-treated supernatant of alveolar macrophage on protein expression of human embryonic lung fibroblasts (MRC-5) by proteomics. Methods The experimental group stimulated MRC-5 with the culture supernatant of rat AM. The control group MRC-5 was treated with an equal amount of AM supernatant without dust. The total MRC-5 protein was extracted for bidirectional coagulation Identify the differentially expressed proteins by gel electrophoresis and perform protein identification by mass spectrometry and database search. Results The number of MRC-5 cells in the control group was (410 ± 21) and that in the dust group was (398 ± 19). The match between the two groups was good. 35 protein spots changed after the dye supernatant was stimulated, and several proteins related to oxidative stress were identified: heat shock protein (HSP27), Per-oxiredoxinⅡ subtype b, calcium network Protein-3 and calreticulin-5 analogs and the like. Conclusion The application of proteomics method can help to screen the differences of protein expression among different treated cells and provide the molecular basis for further study of silicosis mechanism.