RAW264.7细胞泡沫化前后差异蛋白质组分析

Differential Proteomics Research in the RAW264.7 and Foam Cells Induced from RAW264.7

目的应用凝胶内差异显示电泳技术和质谱技术研究小鼠巨噬细胞RAW264.7泡沫化前后蛋白质组的差异。方法体外培养RAW264.7细胞,用氧化型低密度脂蛋白作用使其转变为泡沫细胞。分别提取RAW264.7细胞泡沫化前后的细胞总蛋白,用荧光染料Cy3或Cy5进行标记,与Cy2标记的内标等量混合后在同一胶中进行电泳分离,经不同光激发后扫描得到不同样品的蛋白质组图谱。采用DeCyder6.5软件进行差异蛋白质组分析,筛选出12个差异蛋白质。经质谱鉴定和分析,其中10个蛋白质得到鉴定。结果RAW264.7细胞泡沫化后,应激蛋白70、二硫键异构酶、细胞质肌动蛋白和一种未命名的蛋白质表达量降低,而葡萄糖调节蛋白、烯醇酶、Eno1蛋白、Peroxiredoxin4、Stathmin1和BID蛋白表达量上升。结论本研究建立了巨噬细胞泡沫化前后蛋白质组图谱,并成功进行差异蛋白质组分析,从蛋白质组水平增加了对细胞泡沫化的机制认识,为细胞泡沫化形成动脉粥样斑块的干预研究提供新思路和新靶点。

Aim Two-dimensional difference gel electrophoresis analyse the differential proteins between RAW264.7 cells and foam cells. （2-D DIGE ） and mass spectrum were used to Methods RAW264.7 cells cultured in vitro was transformed to foam cells by treating with ox-LDL. The total proteins of the RAW264.7 cells and foam cells were extracted and labeled With Cy3 and CyS. Each Cy3-1abeled sample and Cy5-labeled sample were mixed on the same 2-D gel along with a Cy2-1abeled mixture of all samples as an internal standard and run on the same gel. All images were analyzed by DeCyder software DIGE technology with internal standard. 12 unique proteins were selected, and in which 10 proteins were successfully identified. Results The expression of Stress protein 70, Disulfide bond isomerase and Cytoplasm actin were decreased in foam cells, then the expression of Glucose regulated protein, Enol protein, Peroxiredoxin 4, Stathmin 1 and BID protein were increased. Conclusions The proteomics maps of RAW264.7 cells and foam cells were established, and some different proteins were selected and identified. Molecular mechanisms about the formation of foam cells was explored from proteomics-level, and provided new routes and new targets for the intervention study of foam cells transformation to atheromatous plaque.