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目的探究硒代蛋氨酸(Selenomethionine,Se-Met)对小鼠巨噬细胞的增殖及巨噬细胞中硒蛋白K(Selenopro-tein K,Sel K)的影响,并观察Sel K在小鼠巨噬细胞中的定位。方法加硒代蛋氨酸(Se-Met)培养巨噬细胞,用CCK-8法检测巨噬细胞的增殖活力,采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹技术检测Se-Met对小鼠巨噬细胞中Sel K m RNA和Sel K蛋白表达的影响;用细胞免疫荧光法观察Sel K在巨噬细胞中的位置。结果 Se-Met能促进巨噬细胞的增殖,其中浓度为1μmol/L Se-Met在处理巨噬细胞24 h和48h后,能显著性地促进巨噬细胞的增殖(P<0.01);1μmol/L Se-Met能使巨噬细胞中Sel K的m RNA含量显著增加(24 h处理组,P<0.05;48 h处理组,P<0.01),另外,Sel K蛋白的含量也有所增加(48 h处理组,P<0.05)。浓度为100μmol/L Se-Met在处理巨噬细胞48 h后,能极显著性地抑制巨噬细胞增殖(P<0.01)。结果还显示Sel K蛋白与内质网标志物KDEL都存在于巨噬细胞内的相同位置上。结论 Se-Met在促进巨噬细胞增殖的同时,使存在于内质网上的Sel K增加。
Objective To investigate the effect of Selenomethionine (Se-Met) on the proliferation of murine macrophages and Selenopro-tein K (Sel K) in macrophages and to observe the effect of selenomethionine on mouse macrophages In the positioning. Methods Macrophages were cultured with Se-Met and the proliferative activity of macrophages was determined by CCK-8 assay. The expression of Se-Met was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting On murine macrophages Sel K m RNA and Sel K protein expression; using immunofluorescence to observe Sel K in macrophages position. Results Se-Met could promote the proliferation of macrophages. The concentration of Se-Met at 1 μmol / L could significantly promote the proliferation of macrophages (P <0.01) after treated with macrophages for 24 h and 48 h (P <0.01) L Se-Met significantly increased the content of sera K m RNA in macrophages (24 h treatment group, P <0.05; 48 h treatment group, P <0.01). In addition, the content of Sel K protein also increased h treatment group, P <0.05). After treated with 100 μmol / L Se-Met for 48 h, Se-Met significantly inhibited the proliferation of macrophages (P <0.01). The results also show that both the Sel K protein and the endoplasmic reticulum marker KDEL are present in the same location within macrophages. Conclusion Se-Met promotes the proliferation of macrophages and increases the Sel K content in the endoplasmic reticulum.