脂多糖刺激脐带间充质干细胞上调 IL-1Ra分泌的研究?

目的：探讨脂多糖(lipopolysaccharide,LPS)激活 Toll 样受体4(Toll-like receptor 4,TLR4)对脐带间充质干细胞(umbilical cord mesenchymal stem cells,UC-MSCs)分泌白介素1受体拮抗剂(interleukin-1 receptor antagonist,IL-1Ra)的影响。方法采用组织块法从脐带华通胶组织中分离培养 UC-MSCs；流式细胞术检测UC-MSCs表面标志物 CD14、CD34、CD29和 CD105的表达；TLR4配体 LPS 以不同剂量和时间刺激 UC-MSCs,采用 qPCR 法和 ELISA 法检测 IL-1Ra 的 mRNA 和蛋白质表达水平。结果原代组织培养7~10 d,组织周边有贴壁细胞爬出,呈长梭形；UC-MSCs 表面标志物 CD29和 CD105呈阳性表达,而 CD14和 CD34呈阴性表达；LPS 刺激 UC-MSCs 后,IL-1Ra 在基因及蛋白质水平的表达均显著增加,差异有统计学意义(P <0.05),并呈剂量与时间依赖性。结论LPS 刺激 UC-MSCs 可上调 IL-1Ra 的分泌。

IL-1 Ra Secretion Up-regulated by Stimulation of LPS in Umbilical Cord Mesenchymal Stem Cells

Objective To explore the impact of IL-1Ra secretion by LPS activating toll-like receptor 4 in umbilical cord mesenchymal stem cells.Methods Human umbilical cord mesenchymal stem cells (UC-MSCs)were isolated and cultured by tissue adherence method from Wharton′s Jelly tissues of umbilical cord.Flow cytometry was performed to assay the surface marker expression of UC-MSCs (CD14, CD34, CD29 and CD105 ). Lipopolysaccharide,a TLR4 ligand,stimulated the UC-MSCs at different dose and time to detect IL-1Ra expression by Quantitative PC-R and ELISA,and detected IL-6 expression as a positive control.Results After primary culture for 7-10 days,UC-MSCs were found around the tissues with spindle shape.Flow cytometry assay showed that UC-MSCs were positive for CD29 and CD105 while negative for CD14 and CD34.mRNA and protein up-regulation of IL-1Ra were dependent on dose and time effect of LPS stimulation in UC-MSCs (P < 0.05 ).Conclusion LPS stimulating umbilical cord mesenchymal stem cells can up-regulate IL-1Ra secretion.