氧化修饰性脂蛋白对C57BL/6J小鼠骨髓源性树突状细胞迁移的影响

Oxidized lipoproteins on the migration of bone marrow derived dendritic cells from C57BL/6J mice

目的:探讨氧化修饰性脂蛋白对C57BL/6J小鼠骨髓源性树突状细胞(bone marrow derived dendritic cells,BM-DCs)迁移的影响。方法:制备C57BL/6J小鼠骨髓细胞悬液,利用树突状细胞专用分离液和细胞粘附性差异去除杂细胞,用rmGM-CSF和rmIL-4使其分化为BMDCs。采用流式细胞术检测BMDCs CD86和MHCⅡ的表达率、免疫荧光技术检测BMDCs CD11c的表达率,以此来鉴定获得细胞为实验所需要的BMDCs。实验分为PBS阴性对照组、LDL组、ox-LDL组、HDL组、oxHDL组和LPS阳性对照组,采用动物实验和Transwell迁移系统分别观察各实验组中BMDCs在体内外的迁移能力。结果:获得的BMDCs CD86、MHCⅡ和CD11c的表达率明显升高,是实验所需的细胞;与相应的脂蛋白组相比,其氧化脂蛋白处理的BMDCs组有更多的细胞发生迁移。结论:氧化脂蛋白可促进C57BL/6J小鼠BMDCs发生迁移。

Objective： To explore the influence of oxidized lipoproteins on the migration of bone marrow derived dendritic cells （BMDCs） from C57BL/6J mice. Methods： The cell suspension was prepared from C57BL/6J mice and the estimated ceils were purified by exclusive use of separating medium for animal dendritic cells and approach of identifying difference of cell adhesion, and the recombinant mouse GM-CSF （rmGM- CSF） and IL-4（rmlL-4） were used to promote monocytes to differentiate and suppress lymphocytes. The BMDCs were estimated by determining the expression BMDCs CD86 and MHCII and BMDCs CD11 c with fluoresoence activated cell sorting（FACS） and i technique, respectively. The experiment was conducted on the groups of chemoattractants of PBS（ negative controls） ,50 μg/nd lipoprotein（ LDL and HDL） , 50 μg/ml oxidized lipoprotein （ oxLDL and oxHDL ） , 1 μg/ml lipopolysaocharide （LPS,positive controls） for observing the migration of BMDCs in vitro and in vivo with Transwell system. Results：The expressions of CD86,MHC Ⅱ and CD11c on the BMDCs were obviously increased, which proved that the BMDCs were the estimated cells. Compared with the groups of lipoprotein ,greater number of ceils in the corresponding oxi- dized lipoprotein groups migrated. Conclusion：Oxidized lipoproteins may promote BMDCs from C57BL/6J mice to migrate.