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目的观察亲环素A(CypA)对THP-1源性泡沫细胞三磷酸腺苷结合盒转运体A1(ABCA1)表达和胆固醇流出的影响及机制。方法 160 nmol/L佛波酯诱导人THP-1单核细胞分化为巨噬细胞,与50 mg/L氧化型低密度脂蛋白(ox-LDL)共孵育,使其荷脂形成泡沫细胞,常规体外培养细胞;实验分为对照组和Cyp A处理组;液体闪烁计数仪检测细胞胆固醇流出水平,高效液相色谱检测细胞内脂质成分,实时荧光定量PCR和Western blot检测ABCA1表达,Western blot检测核因子κB(NF-κB)核转位水平;NF-κB抑制剂小白菊内酯抑制NF-κB活化;脂质体2000转染CD147 siRNA至THP-1源性泡沫细胞。结果 CypA显著抑制THP-1源性泡沫细胞胆固醇流出,促进NF-κB核转位,下调ABCA1表达;NF-κB抑制剂小白菊内酯拮抗CypA对ABCA1表达及胆固醇流出的抑制作用;用siRNA干扰CD147后,显著抑制Cyp A诱导的NF-κB核转位,上调ABCA1表达,促进胆固醇流出。结论 CypA可能通过CD147激活NF-κB,下调ABCA1表达,抑制THP-1源性泡沫细胞胆固醇流出。
Objective To investigate the effect and mechanism of cyclophilin A (CypA) on THP-1-derived foam cell adenosine triphosphate binding cassette transporter A1 (ABCA1) expression and cholesterol efflux. METHODS: THP-1 monocytes were induced to differentiate into macrophages by phorbol ester at 160 nmol / L and incubated with ox-LDL (50 mg / L) to form foam cells. Conventional The cells were cultured in vitro. The experiment was divided into control group and Cyp A treatment group. The efflux of cholesterol was detected by liquid scintillation counter. The lipid composition was detected by high performance liquid chromatography. The expression of ABCA1 was detected by real-time fluorescence quantitative PCR and Western blot. Nuclear factor kappa B (NF-κB) nuclear translocation; NF-κB inhibitor Parthenolide inhibited NF-κB activation; lipofectamine 2000 transfected CD147 siRNA into THP-1-derived foam cells. Results CypA significantly inhibited the efflux of cholesterol from THP-1-derived foam cells, promoted the nuclear translocation of NF-κB and down-regulated the expression of ABCA1. Inhibition of CypA by NF-κB inhibitor CypA played an inhibitory role on the expression of ABCA1 and cholesterol efflux. CD147 significantly inhibited Cyp A-induced nuclear translocation of NF-κB, up-regulated ABCA1 expression and promoted cholesterol efflux. Conclusion CypA may activate NF-κB through CD147, down-regulate the expression of ABCA1 and inhibit the efflux of THP-1-derived foam cells.