炎症环境中GM-CSF对骨髓干细胞迁移能力及炎症因子的影响

Effects of GM-CSF on migration of bone marrow stem cells and inflammatory factors under inflammatory environment

目的探讨炎症环境下粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)对骨髓间充质干细胞(mesenchymal stem cells,MSCs)迁移能力及炎症因子的影响。方法将MSCs细胞与RAW264.7细胞采用Transwell共培养,分为对照组(正常培养液)、LPS组(1μg/ml LPS)和LPS+GM-CSF组(20μg/L GM-CSF与1μg/ml LPS共处理)。Transwell小室检测MSCs细胞迁移能力。ELISA检测RAW264.7细胞上清中TNF-α、COX-2、iNOS和IFN-γ水平。qPCR检测MSCs细胞CSF、IL-6、IL^(-1)1和IL-7的表达。结果与对照组相比,LPS组MSCs迁移细胞数量显著增多(P<0.05),CSF表达显著下调(P<0.05),IL-6、IL^(-1)1和IL-7表达显著上调(P<0.05),RAW264.7细胞上清中TNF-α、COX-2、iNOS和IFN-γ水平显著升高(P<0.05)。与LPS组相比,LPS+GM-CSF组MSCs迁移细胞数量显著增多(P<0.05),CSF表达显著上调(P<0.05),IL-6、IL^(-1)1和IL-7表达显著下调(P<0.05),RAW264.7细胞上清中TNF-α、COX-2、iNOS和IFN-γ水平显著降低(P<0.05)。结论GM-CSF可增强炎症环境下MSCs细胞迁移能力并缓解炎症反应。

Objective To investigate the effect of granulocyte-macrophage colony-stimulating factor(GM-CSF)on the migration of bone marrow mesenchymal stem cells(MSCs)and the inflammatory factors under the inflammatory environment.Methods MSCs cells and RAW264.7 cells were co-cultured in Transwell,and divided into normal control group(culture medium),LPS group(1μg/ml LPS)and LPS+GM-CSF group(20μg/L GM-CSF and 1μg/ml LPS co-treatment).The migration ability of MSCs was detected by Transwell assay,and the levels of TNF-α,COX-2,iNOS and IFN-γin the supernatant of RAW264.7 cells were detected by ELISA.The expression levels of CSF,IL-6,IL^(-1)1 and IL-7 in MSCs were detected by qPCR.Results Compared with control group,the migration of MSCs in LPS group was increased significantly(P<0.05),the expression of CSF was significantly down-regulated(P<0.05),the expression levels of IL-6,IL^(-1)1 and IL-7 were significantly up-regulated(P<0.05),and the levels of TNF-α,COX-2,iNOS and IFN-γin the supernatant of RAW264.7 cells were significantly increased(P<0.05).Compared with LPS group,the migration of MSCs was significantly increased in LPS+GM-CSF group(P<0.05),the expression of CSF was significantly up-regulated(P<0.05),the expression levels of IL-6,IL^(-1)1 and IL-7 were significantly down-regulated(P<0.05),and the levels of TNF-α,COX-2,iNOS and IFN-γin the supernatant of RAW264.7 cells were significantly decreased(P<0.05).Conclusion GM-CSF can enhance the cell migration ability of MSCs in inflammatory environment and alleviate the inflammatory response.