硒代蛋氨酸对小鼠巨噬细胞增殖及硒蛋白K的影响

Effect of Se-Met on mouse macrophage proliferation and its intracellular protein-Sel K

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 explore the effect of selenomethionine （Se-Met）on mouse macrophage proliferation and itsintracellular protein-selenoprotein K（SelK ）,and detect the localization of SelK in mouse macrophage. Methods Theproliferation of macrophage were measured by CCK-8 assay after treatment of Se-Met.The SelK mRNA and protein weredetected by RT-PCR and Western blot respectively. The localization of SelK protein in mouse macrophage was determined byimmunofluorescence technique. Results Se-Met could promote proliferation of macrophage.The proliferation of mousemacrophage was significantly 24 h and 48 h after treatment with Se-Met at the concentration of 1μmol/L （P〈 0.01）respectively. The proliferation of mouse macrophage was significantly inhibited 48h after treatment with Se-Met at theconcentration of 100 μmol/L （P〈0.01）.Se-Met at the concentration of 1 μmol/L increased the SelK mRNA level inmacrophages after treating for 24 h （P〈0.05） and 48 h （P〈0.01） respectively. Moreover, SelK protein level also increased aftertreatment for 48 h （P 〈0.05）. Immunofluorescence showed that SelK protein and endoplasmic reticulum markers （KDEL）existed in the same place in macrophage. Conclusion Se-Met can promote the proliferation of macrophage, meanwhile itcan also make the increase of SelK level.