论文部分内容阅读
目的探讨硒蛋白K(Selenoprotein K,SelK)基因沉默(RNA silence,RNAi)对小鼠巨噬细胞增殖、吞噬和迁移的影响。方法制作并包装小鼠巨噬细胞SelK慢病毒干扰株,将细胞分为未转染正常对照组、空载体慢病毒转染组和慢病毒干扰组。建立稳定干扰的巨噬细胞株后,应用实时定量RT-PCR法和蛋白质免疫印迹法检测SelK的干扰效率,分别应用CCK8法、巨噬细胞吞噬鸡红细胞实验和Transwell法检测细胞增殖、细胞吞噬能力和细胞迁移能力。结果与空载体慢病毒转染组相比,SelK慢病毒干扰组巨噬细胞的mRNA和SelK水平显著降低,差异均有统计学意义(P<0.05);48 h后,与空载体慢病毒转染组相比,慢病毒干扰组巨噬细胞的增殖受到了明显的抑制,差异有统计学意义(P<0.01),同时慢病毒干扰组巨噬细胞穿过小室的细胞数显著低于空载体慢病毒转染组(P<0.05),慢病毒干扰组巨噬细胞能吞噬鸡红细胞的巨噬细胞数量显著较少。结论 Selk基因沉默能使小鼠巨噬细胞的增殖、吞噬和迁移能力降低,从而改变其抗感染免疫能力。
Objective To investigate the effect of selenoprotein K (SelK) gene silencing on the proliferation, phagocytosis and migration of murine macrophages. Methods The murine macrophage SelK lentiviral vector was prepared and packaged. The cells were divided into untransfected normal control group, empty vector lentivirus transfection group and lentiviral interference group. After the establishment of stable macrophage cell line, real-time quantitative RT-PCR and Western blotting were used to detect the interference efficiency of SelK. CCK8 assay, macrophage phagocytosis chicken erythrocyte assay and Transwell assay were used to detect the cell proliferation and phagocytosis And cell migration ability. Results Compared with empty vector lentiviral transfection group, SelK lentiviral interference group mRNA and SelK levels of macrophages were significantly lower (P <0.05); after 48 h, compared with empty vector lentiviral transfection group Compared with the control group, the proliferation of macrophages in lentiviral interference group was significantly inhibited (P <0.01), while the number of macrophages passing through the cell in lentiviral interference group was significantly lower than that in empty vector Lentiviral transfection group (P <0.05), lentiviral interference group of macrophages can swallow chicken red blood cells significantly fewer macrophages. Conclusion Silencing of Selk gene can decrease the ability of mouse macrophages to proliferate, phagocytize and migrate, thereby changing their anti-infective ability.