摘要
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 observe the change of protein profile in human embryonic pulmonary fibroblasts caused by supernatant fluid of silica-treated alveolar macrophage.Methods MRC-5 cell lines were stimulated by supernate of rat alveolar macrophage（AM） treated with silica in the experiment group and without silica in the control group.After 24 hours of cultivation,MRC-5 proteins were extracted for 2D Gel Electrophoresis analysis and identified by MS.Results There were 410±21 and 398±19 protein spots detected in experiment group and in control group, respectively. The match of the expression of cellular proteins between groups was relatively good. Proteins in 35 spots were significantly observed to be differential expressed, and several proteins related to oxidative stress were identified to he HSP27, peroxiredoxin Ⅱ-b, similar to calreticulin isoform 3 and similar to calreficulin isofonn 5. Conclusion The utilization of proteomic technology is effective in screening different protein expressions and can provide molecular targets for the study of silicosis mechanism.
出处
《中国职业医学》
CAS
北大核心
2009年第4期281-283,共3页
China Occupational Medicine
基金
广西青年科学基金资助(桂科青0832047)
广西大型仪器协作共用网资助
