干扰SelK对小鼠巨噬细胞非特异性免疫应答的影响

Effect of Sel K gene silencing on proliferation, phagocytosis and migration of mouse macrophage

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 observe the effect of selenoprotein K（SelK） gene silencing on mouse macrophage proliferation, phagocytosis and migration. Methods The lentiviral RNA interferenced vector of mouse selk gene was constructed.The cells were divided into control group,empty vector group and RNAi vector group.The selenoprotein K （SelK） mRNA and protein were detect by RT-PCR and Western blot respectively.CCK-8 assay was used to detect cell proliferation, macrophages swallowing chicken erythrocytes assay was used to detect cell phagocytosis and transwell was used to detect cell migration. Results The expression of SelK mRNA and protein in the RNAi vector group was decreased compared with the empty vector group showed significant decrease （P〈0.05）. The proliferation of mouse macrophages was significantly inhibited 48 hours after SelK gene was silenced （P〈0.01）. and the ability of phagocytosis and migration was decreased as well compared with that in control group （P〈0.05）. Conclusion SelK gene silencing could inhibit proliferation,phagocytosis and migration of mouse macrophage.