炎症环境下巨噬细胞对小鼠骨髓间质干细胞迁移能力的影响  

Effect of macrophages on the migration of mouse bone marrow mesenchymal stem cells in inflammation

作　　者：高硕[1] 蔡梦洁[1] 毛飞[1] 杨婷婷[1] 张洁[1] 张玲[1] 赵露露[1] 坎吉克孜.阿不来提 钱晖[1] 许文荣[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2013年第3期201-206,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金重大研究计划资助项目(91129718);国家自然科学基金资助项目(81000181);江苏省高校自然科学基金资助项目(10KJB310002);江苏大学高级人才启动基金资助项目(10JDG094);江苏大学大学生科研立项课题(11A387)

摘　　要：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＆lt;0.01）,while RAW264.7 cells remarkably increased the migratory ability of MSCs under the stimulation of LPS（P＆lt;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＆lt;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.

关 键 词：骨髓间质干细胞 巨噬细胞 细胞迁移 细胞因子 炎症 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] Q291[医药卫生—基础医学]