目的:探讨脂多糖(LPS)存在的炎症环境下巨噬细胞株RAW264.7对小鼠骨髓间质干细胞(mesenchymalstem cells,MSCs)的基因表达及迁移能力的影响。方法:采用全骨髓贴壁法分离小鼠骨髓MSCs,与RAW26 4.7细胞进行Transwell迁移及共培养实验,分为培养液对照组,RAW264.7作用组,LPS作用组及RAW264.7与LPS共作用组。Transwell迁移实验比较各组MSCs的迁移数量,同时qRT-PCR检测各组MSCs表达转化生长因子-β1(TGF-β1)、白介素-6(IL-6)和单核细胞趋化蛋白-1(MCP-1)mRNA水平。结果:LPS刺激巨噬细胞活化,使其高表达IL-6、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)及MCP-1。RAW264.7作用组MSCs迁移数量多于对照组(P〈0.01)。与LPS作用组相比,LPS活化的RAW264.7显著增加MSCs的迁移数量(P〈0.01)。qRT-PCR结果显示活化的RAW264.7增加了MSCs的TGF-β1、IL-6和MCP-1 mRNA表达(P〈0.05)。结论:炎性条件下巨噬细胞能增强MSCs的迁移能力并改变其细胞因子的表达水平。
Objective: To investigate the effect of macrophage cell line RAW264.7 on the migration and expression of TGF-β1,IL-6 and MCP-1 gene level of mouse bone marrow derived mesenchymal stem cells(MSCs) under the stimulation of lipopolysaccharide(LPS).Methods: Adherent method was used to isolate mouse bone marrow mesenchymal stem cells.MSCs were co-cultured with RAW264.7 through transwell assay.Co-culture experiments were divided into four groups: culture medium control,RAW264.7 cells,LPS and RAW264.7 cells stimulated by LPS.Cell transwell migration assay were performed to detect the number of migrated MSCs under the four different conditions.At the same time,qRT-PCR was performed for the quantification of TGF-β1,IL-6 and MCP-1genes.Results: RAW264.7 expressed high level of TNF-α,IL-6,IL-1β and MCP-1 gene by LPS stimulation.In the presence of RAW264.7,the migrated MSCs numbers were higher than controls(P<0.01),while RAW264.7 cells remarkably increased the migratory ability of MSCs under the stimulation of LPS(P<0.01).Meanwhile,the expression of TGF-β1,IL-6 and MCP-1 genes were enhanced by co-culture with RAW264.7 in the presence of LPS(P<0.05).Conclusion: Macrophage cell line RAW264.7 enhanced the migratory ability and exchanged the cytokine expression of MSCs under the stimulation of LPS,which provided mechanistic insights into the recruitment of MSCs in inflammation.