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目的探讨分枝菌酸(MA)包被聚苯乙烯微球诱导小鼠RAW264.7巨噬细胞泡沫细胞化的条件及特性。方法将RAW264.7细胞分为空白对照组和分别包被(0、25、50、75、100)μg/mL MA的聚苯乙烯微球实验组,分别将细胞与聚苯乙烯微球混匀、共同孵育24、48、72 h、5 d和8 d后,油红O染色后显微镜下观察细胞形态,采用总胆固醇试剂盒和游离胆固醇试剂盒测定各组泡沫细胞内胆固醇酯的含量,采用MTT法检测细胞增殖水平。结果当包被MA浓度为75μg/mL、吞噬时间为24 h时,RAW264.7能形成典型的泡沫细胞,其细胞内胆固醇酯相对含量为60.98%±1.32%;随着吞噬时间延长,形成泡沫细胞所需的MA的浓度逐渐递减;且随着细胞泡沫化程度增加,细胞增殖活力逐渐减弱。结论 MA包被聚苯乙烯微球可快速诱导巨噬细胞形成泡沫细胞,MA浓度与细胞泡沫化程度呈正相关,且有时间依赖性。
Objective To investigate the conditions and characteristics of foam cell maturation induced by mycobacterium acid (MA) coated polystyrene microspheres in RAW264.7 macrophages. Methods RAW264.7 cells were divided into blank control group and polystyrene microspheres coated with 0, 25, 50, 75, 100 μg / mL MA respectively. The cells were mixed with polystyrene microspheres , And incubated for 24, 48, 72, 5 and 8 days. The morphology of the cells was observed under oil red O staining and the content of cholesterol ester in foam cells was determined by total cholesterol kit and free cholesterol kit. Cell proliferation was detected by MTT assay. Results When the concentration of coating MA was 75μg / mL and phagocytosis time was 24 h, the typical foam cells were formed in RAW264.7 with the relative content of intracellular cholesterol ester of 60.98% ± 1.32%. With the phagocytic time prolonged, foam formed The concentration of MA required for cells gradually decreased; and as the degree of cell foaming increased, cell proliferation activity gradually decreased. Conclusion MA coated polystyrene microspheres can rapidly induce the formation of foam cells in macrophages. There is a positive correlation between the concentration of MA and the degree of cellular foaming in a time-dependent manner.