摘要
目的探讨雄激素二氢睾酮(DHT)对雌性巨噬细胞系J774.1细胞的植物血凝素样氧化低密度脂蛋白受体1(LOX-1)表达及泡沫细胞形成的影响。方法体外培养J774.1细胞,通过氧化低密度脂蛋白(OX—LDL)诱导巨噬细胞向泡沫细胞转化,采用1×10^-9mol/L和1×10^-8mol/L浓度的DHT进行预处理,提取细胞mRNA及蛋白行实时定量PCR及Western印迹法检测LOX-1mRNA及蛋白表达,油红O染色观察泡沫细胞形态,按照泡沫细胞数/视野及每个视野中油红染色的颗粒面积进行定量分析。结果与对照组(加入乙醇)相比,ox—LDL明显增加J774.1细胞的LOX-1mRNA(4.71±0.31比1.00±0.06)及蛋白(1.75±0.11比1.04±0.04)相对表达水平(均P〈0.01)。应用1×10^-9mol/L和1×10^-9mol/L浓度的DHT可以降低ox-LDL诱导的LOX-1mRNA(2.81±0.46、2.29±0.21比4.71±0.31,均P〈0.05)和蛋白表达水平(1.35±0.06、1.09±0.04比1.75±0.11,均P〈0.05)。这种作用可通过雄激素受体(AR)阻滞剂逆转87.6%(P=0.004)。油红O染色表明ox—LDL明显增加泡沫细胞形成(45.9±3.7比0.0±0.0,P〈0.01)。应用1×10^-9mol/L和1×10^-8mol/L浓度的DHT可以抑制ox-LDL诱导的泡沫细胞形成(泡沫细胞数/视野定量分析:36.0±3.0、29.1±1.3比45.9±3.7,均P〈0.05;油红O相对染色面积定量分析:7983±1035、4060±390比14750±2489,均P〈0.05)。结论1×10^-9mol/L和1×10^-8mol/L浓度的DHT通过AR可以减少雌性巨噬细胞系J774.1细胞的LOX-1表达,并抑制J774.1细胞向泡沫细胞转化。
Objective To investigate the effect of dihydrotestosterone (DHT) on lectin-like ox- low-density lipoprotein (LDL) receptor (LOX-1)expression and foam cell formation in the female macrophage cell line J774. 1. Methods In cultured J774. 1 cells, after pretreated with DHT at concentrations of 1×10^-9 mol/L and 1 ×10^-8 mol/L, ox-LDL-induced LOX-1 expression and foam cell formation were investigated by quantitative real-time PCR, Western blotting, and oil-red O staining. Results DHT at concentrations of 1×10^-9 mol/L and 1×10^-8 mol/L inhibited ox-LDL-induced LOX-1 mRNA (2. 81 ± 0.46 and 2. 29 ± 0. 21 vs 4. 71 ± 0. 31, both P 〈 0. 01 ) and protein expression ( 1.35 ±0. 06 and 1.09 ± 0. 04 vs 1.75 ± 0. 11, both P 〈 0. 05 ). The effect was partly reversed by the androgen receptor (AR) blocker flutamide (87. 6%, P = 0. 004). Oil-red O staining also revealed that DHT at concentrations of 1 ×10^-9 mol/L and 1 ×10^-8 mol/L suppressed ox-LDL-induced foam cell formation as quantified by the number of foam cells per high-power field (HPF) ( 36.0 ± 3.0 and 29. 1± 1.3 vs 45.9 ± 3.7, both P 〈 0.05 ) and by the area of oil-red O stained particles per HPF (7 983 ± 1 035 and 4 060 ± 390 vs 14 750 ± 2 489, both P 〈 0. 05 ). Conclusion DHT at concentrations of 1 ×10^-9 mol/L and 1 ×10^-8 mol/L decreases LOX-1 expression and foam cell formation via AR.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2016年第42期3403-3407,共5页
National Medical Journal of China
基金
国家自然科学基金(81170779)