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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 gene expression and migration of mouse bone marrow mesenchymal stem cells (MSCs) in the presence of lipopolysaccharide (LPS). Methods: Bone marrow MSCs were isolated by whole bone marrow adherent method and cultured with RAW26.7 cells for transwell migration and co-culture. The cells were divided into culture medium control group, RAW264.7-treated group, LPS-treated group and RAW264.7 with LPS group. Transwell migration assay was used to compare the migration of MSCs in each group. Meanwhile, the expressions of TGF-β1, IL-6 and MCP-1 were detected by qRT-PCR. -1) mRNA levels. RESULTS: LPS stimulated the activation of macrophages, which resulted in the high expression of IL-6, TNF-α, IL-1β and MCP-1. The number of MSCs migrated in RAW264.7 group was more than that in control group (P <0.01). Compared with LPS group, LPS-activated RAW264.7 significantly increased the number of MSCs migrated (P <0.01). qRT-PCR results showed that activated RAW264.7 increased the mRNA expression of TGF-β1, IL-6 and MCP-1 in MSCs (P <0.05). Conclusion: Macrophages can enhance the migration of MSCs and change the expression of cytokines in inflammatory conditions.