氧化修饰低密度脂蛋白对C57BL/6J小鼠骨髓源性树突状细胞成熟和迁移的影响

Oxidized low density lipoprotein promotes the maturation and migration of bone marrow derived dendritic cells from C57BL/6J mice

目的:探讨氧化修饰低密度脂蛋白(oxidized low density lipoprotein,oxLDL)对C57BL/6J小鼠骨髓源性树突状细胞(bone marrow derived dendritic cells,BMDCs)迁移、成熟活化及部分免疫功能的影响。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黏附性差异去除杂细胞,rmGM-CSF和rmIL-4使其分化为BMDCs。实验分为PBS阴性对照组、LDL组、oxLDL组和LPS阳性对照组。采用流式细胞术检测BMDCsCD86和MHC Ⅱ的表达率;液体闪烁计数检测各实验组BMDCs刺激同源T淋巴细胞增殖的能力;ELISA法检测分泌到培养基中的IL-12和IL-10,并借助Transwell迁移系统,观察oxLDL对BMDCs迁移的影响。结果:与PBS组相比,oxLDL处理的BMDCs,其CD86和MHC Ⅱ表达率、混合淋巴细胞反应和细胞因子分泌都明显升高;但仍低于LPS组。同样与PBS组相比,oxLDL组迁移到Transwell下室面的细胞数目明显增加,大约是LDL组的3倍,但仍低于LPS组(P<0.05)。结论:oxLDL可促进BMDCs成熟,并趋化BMDCs发生迁移。

Objective To investigate the effects of oxidized low density lipoprotein （oxLDL） on the migration, maturation and immune functions of the bone marrow derived dendritic cells （BMDCs）from C57BL/6J mice. Methods The cell suspension was made from bone marrow of C57BL/6J mice and purified by the strategy of differences in cell adhesion using separating medium for animals. The rmGM-CSF and rmIL-4 were used to promote monocytes differentiate into BMDCs. The oxLDL（50 ug/mL） was used to induce BMDCs, with LDL（50 ug/mL） as homologous protein controls, PBS and LPS （ 1 ug/mL） as negative and positive controls. The expressions of CD86 and MHC Ⅱ were detected by fluorescence-activated cell sorting （FACS）, the ability of BMDCs to stimulate homologous T cells proliferation was tested by liquid scintillation counting （LSC）, the expressions of cytokine IL-12 and IL-10 were detected by enzyme-linked immunosorbent assay （ELISA）. The migration of BMDCs were observed using the transwell system. Result Compared with the PBS group, the expressions of CD86 and MHC Ⅱ , the cpm of mixed lymphocyte reaction （MLR）, and the secretion of IL-12 and IL-10 were all increased in the oxLDL group. The number of migrating cells was larger in the oxLDL group than that of the LDL group, but smaller than that of LPS group（P 〈 0.05）. Conclusion The OxLDL can promote the maturation and migration of BMDCs in vitro.