摘要
目的探讨氧化型高密度脂蛋白对C57BL/6J小鼠骨髓源性树突状细胞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粘附性差异去除杂细胞,用重组小鼠粒细胞-巨噬细胞集落刺激因子和重组小鼠白细胞介素4使其分化为骨髓源性树突状细胞。实验分为PBS阴性对照、高密度脂蛋白组、氧化型高密度脂蛋白组和脂多糖阳性对照组。采用流式细胞术检测骨髓源性树突状细胞CD86和MHCⅡ的表达率;液体闪烁计数检测各实验组骨髓源性树突状细胞刺激同源T淋巴细胞增殖的能力;ELISA法检测分泌到培养基中的白细胞介素12和白细胞介素10。结果与PBS组相比,氧化型高密度脂蛋白处理的骨髓源性树突状细胞,其CD86、MHCⅡ表达率、混合淋巴细胞反应液体闪烁计数和细胞因子分泌量都明显升高;高密度脂蛋白组各项指标升高不如氧化型高密度脂蛋白明显。结论氧化型高密度脂蛋白可促进C57BL/6J小鼠骨髓源性树突状细胞成熟活化和细胞因子的分泌。
Aim To explore the influence of oxidized high density lipoprotein （ox-HDL） on the maturation and some immune functions of bone marrow derived dendritic cells （ BMDC） from C57BL/6J mice. Methods The cell suspension was made of C57BL/6J mice, purified by use of separating medium for animals and the difference of cell adhesion, and recombinant mouse GM-CSF （ rmGM-CSF ） and recombinant mouse IL-4 were used to promote monocytes to differentiate and to suppress lymphocytes. 50 mg/L ox-HDL was added to stimulate BMDC, using 50 mg/L HDL as homologous protein control, PBS as negative control and 1 mg/L lipopolysaccharide （LPS） as positive control. The CD86 and MHC Ⅱ expressing rates were detected with fluorescence-activated cell sorting （FACS）. Liquid scintillation counting （ LSC ） was used in mixed lymphocyte reactions （MLR） to reflect the ability of BMDC to stimulate homologous T cells proliferation. Cytokines IL-12 and IL-10 were detected by ELISA. Result Compared with PBS group, ox-HDL group increased the expressions of CD86 and MHC Ⅱ, counts per minute （ cpm） of MLRs, secretion of IL-12 and partly IL-10. The increment of HDL group was not remarkable. Conclusion ox-HDL could promote BMDC to mature in vitro.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2009年第1期39-42,共4页
Chinese Journal of Arteriosclerosis
基金
湖南省教育厅项目(编号06C692)
