胚胎干细胞来源神经干细胞调控巨噬细胞的实验研究

Effect of embryonic stem cells-derived neural stem cells on macrophage in vitro

目的:观察胚胎干细胞(Embryonic stem cell,ESC)来源的神经干细胞(Neural stem cells,NSCs)对骨髓巨噬细胞的增殖、吞噬及分泌细胞因子的影响,为探讨NSCs免疫调节及在重大疾病中的应用做铺垫。方法:培养骨髓巨噬细胞,收集胚胎干细胞来源NSCs(ESC-NSCs)的上清液处理巨噬细胞,然后利用磺酰罗丹明B(Sulforhodamine B,SRB)法检测增殖情况;利用红荧光beads检测吞噬能力;ELISA法检各组TNF-α和IL-1β表达情况。结果:成功从ESC获得具有NSCs特征的细胞。对照组、NSC组巨噬细胞增殖率分别为100%、(126.29±5.41)%,beads吞噬率分别为(70.23±2.57)%、(90.32±8.49)%。相比对照组,NSC组巨噬细胞TNF-α和IL-1β含量下降(P<0.05)。结论:ESC来源的NSCs具有促进骨髓巨噬细胞增殖及增强其吞噬能力,并抑制其炎性因子分泌,为NSCs免疫调节作用提供新的依据。

Objective： To explore the influence of embryonic stem cells-derived neural stem cells on the proliferation and secretion cytokines of bone marrow-derived macrophages. Methods： Mouse bone marrow derived macrophages were isolated and cultured in L929 medium. After macrophages were treated with NSCs supernatant for 3 days,SRB method was used to detect the proliferation of macrophages. The phagocytosis of macrophages were detected by incubating with RFP-Beads for 1 h. Meanwhile,the expression of TNF-α and IL-1β were detected by ELISA. Results： NSCs were successfully induced from ESC. In control group and NSC group,the proliferation rate of macrophages were 100 % and（ 126. 29 ± 5. 41） %,the phagocytosis rate were（ 70. 23 ± 2. 57） % and（ 90. 32 ±8. 49） %. Compared to the control group,the levels of IL-6,IL-1β in macrophage treated with NSCs decreased（ P〈0. 05）. Conclusion：ESC-derived NSCs can promote the proliferation and phagocytosis of bone marrow-derived macrophage,and suppress the secretion of pro-inflammatory cytokines.