目的研究核因子-κB(NF-κB)抑制剂二硫代氨基甲酸吡咯烷(PDTC)对THP-1巨噬细胞脂质蓄积和胆固醇流出的影响。方法 60 nmol/L PMA孵育THP-1巨噬细胞24 h后,分为3组:对照组、氧化型低密度脂蛋白(ox-LDL)组(100μg/mL)和PDTC组(100μg/mL ox-LDL+50μmol/L PDTC)。用油红O染色观察细胞内脂质蓄积情况,高效液相色谱分析法检测细胞内胆固醇水平,用液体闪烁计数法观察细胞内胆固醇的流出。结果PDTC抑制ox-LDL诱导的THP-1巨噬细胞脂质蓄积,明显降低细胞内总胆固醇、游离胆固醇和胆固醇酯的含量,增加载脂蛋白A-Ⅰ(apoA-Ⅰ)介导的胆固醇流出,但对HDL介导的胆固醇流出无明显影响。结论 PDTC抑制ox-LDL诱导的THP-1巨噬细胞脂质蓄积,促进apoA-Ⅰ介导的胆固醇流出。
Objective To investigate the effect of NF-κB inhibitor pyrrolidinedithiocarbamate(PDTC) on lipid accumulation and cholesterol efflux in THP-1 macrophages induced by ox-LDL.Methods Human THP-1 macrophage was induced using phorbol-12-myristate acetate(PMA,160 nmol/L) for 24 h.Then cells were divided into three groups:control group,ox-LDL group(100 μg/mL) and PDTC group(100 μg/mL ox-LDL+50 μmol/L PDTC).Oil red O staining was used to observe the intracellular lipid droplets.Cellular cholesterol was determined by high performance liquid chromatography analysis.Cholesterol efflux was determined by liquid scintillation counting.Results PDTC inhibited the lipid accumulation in THP-1 macrophage induced by ox-LDL.Cellular total cholesterol,free cholesterol and cholesterol ester were decreased by application of PDTC.PDTC increased apoA-Ⅰ mediated cholesterol efflux and had no significant effect on HDL-mediated cholesterol efflux.Conclusion PDTC inhibits lipid accumulation induced by ox-LDL and increases apoA-Ⅰ mediated cholesterol efflux in THP-1 macrophage.