荧光标记结合超声裂解测定泡沫细胞胆固醇流出

Determination of Cholesterol Efflux from Foam Cells Using Fluorescence Labeled Cholesterol with Ultrasonic Lysis

利用荧光基团NBD标记的胆固醇(NBD-胆固醇)并结合超声裂解检测THP-1源性巨噬泡沫细胞胆固醇流出率。佛波酯(PMA)刺激人单核THP-1细胞分化为巨噬细胞,NBD-胆固醇作用于乙酰化低密度脂蛋白(ac-LDL)诱导的巨噬细胞源性泡沫细胞后,测定载脂蛋白A-1(apoA-1)介导的胆固醇流出率。结果显示,与对照ac-LDL组相比,apoA-1介导的泡沫细胞胆固醇流出率增加了78.57%(P<0.01)。[3H]-标记胆固醇处理的泡沫细胞,apoA-1介导的胆固醇流出率与对照组相比,增加了80.12%(P<0.001)。NBD-标记胆固醇与超声裂解细胞相结合的方法,可作用一种替代同位素标记胆固醇的方法对泡沫细胞胆固醇流出进行检测。

To determine the cholesterol efflux rate of THP-1-derived macrophage foam cells by using the fluorescent group NBD-labeled cholesterol(NBD-cholesterol)with ultrasonic lysis method.Human THP-1 cells were differentiated into macrophages with phorbol-myristate-13-acetate(PMA),and foam cells induced with acetylated low density lipoprotein(ac-LDL)were treated with NBD-cholesterol,and then apolipoprotein A-1(apoA-1)-mediated cholesterol efflux was measured.The result showed that the apoA-1-mediated cholesterol efflux rate was increased by 78.57%(P<0.01),to compared with the control ac-LDL group.In[3H]-labeled foam cells,the apoA-1-mediated cholesterol efflux rate increased by 80.12%compared to the control cells(P<0.001).The combination of NBD-labeled cholesterol with ultrasonic lysis may be an alternative method to detect cholesterol efflux from foam cells.