阿托伐他汀对THP-1巨噬细胞脂质蓄积及CD36表达的影响

Effect of Atorvastatin on Lipid Accumulation and CD36 Expression in THP-1 Macrophages

目的观察阿托伐他汀对氧化型低密度脂蛋白诱导的THP-1巨噬细胞脂质蓄积及CD36表达的影响。方法用50mg/L氧化型低密度脂蛋白与不同浓度的阿托伐他汀(0、0.312、1.25和5μmol/L)共同孵育THP-1巨噬细胞24h,以空白组作对照,用液体闪烁计数法检测细胞[3H]胆固醇流入情况,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总胆固醇水平,逆转录聚合酶链反应与免疫印迹分析法分别检测THP-1巨噬细胞CD36mRNA和蛋白的表达。结果阿托伐他汀使THP-1巨噬细胞胆固醇流入减少,对照组胆固醇流入为35.90%±2.36%,0、0.312、1.25和5μmol/L阿托伐他汀组胆固醇流入分别为47.10%±3.18%、41.20%±2.88%、35.10%±2.35%和28.30%±1.98%;阿托伐他汀能抑制THP-1巨噬细胞对氧化型低密度脂蛋白的摄取,油红O染色可见氧化型低密度脂蛋白+阿托伐他汀组细胞内脂滴较氧化型低密度脂蛋白组明显减少,且脂滴颗粒体积变小;高效液相色谱分析发现,对照组细胞总胆固醇含量为78.24±11.35mg/g,0、0.312、1.25和5μmol/L阿托伐他汀组细胞总胆固醇含量分别为123.13±15.92mg/g、115.36±13.18mg/g、107.52±12.05mg/g和98.03±10.24mg/g。阿托伐他汀使氧化型低密度脂蛋白诱导的THP-1巨噬细胞CD36mRNA和蛋白的表达下调。结论阿托伐他汀引起氧化型低密度脂蛋白诱导的THP-1巨噬细胞CD36表达下调,并使脂质蓄积减少。

Aim To investigate the effect of atorvastatin on oxidized low density lipoprotein （ox-LDL）-indueed lipid accumulation and CD36 expression in THP-1 maerophages. Methods THP-1 maerophages were co-incubated with 50 mg/L ox- LDL and different concentrations of atorvastatin （0, 0.312, 1.25 and 5μmol/L） for 24 h, and the unmixed THP-1 macrophages culture was used as the control group. [^3H]-labeled Cholesterol influx was determined by FJ-2107P type liquid scintillator. Oil red O staining was used to observe the intracellular lipid droplets. Cellular total cholesterol was determined by high performance liquid chromatography analysis. CD36 mRNA and protein levels were determined by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting respectively. Results Atorvastatin decreased CD36 in both protein and mRNA levels. Atorvastatin decreased cholesterol influx in THP-1 macrophages. The levels of cholesterol influx were 35.90% ± 2.36% （control）, 47.10%±3.18% （0 μmol/L atorvastafin）, 41.20%±2.88% （0.312 μmol/L atorvastatin）, 35.10%±2.35% （1.25 μmol/L atorvastatin）, 28.30% ± 1.98% （5 μmol/L atorvastatin） respectively. Oil red O staining showed intracellular lipid droplets were minor with the presence of atorvastatin. High performance liquid chromatography analysis demonstrated the amount of cellular total cholesterol were 78.24 ± 11.35 mg/g （control）, 123.13 ± 15.92 mg/g （0 μmol/L atorvastatin）, 115.36 ± 13.18 mg/g （0.312 μmol/L atorvastafin）, 107.52 ± 12.05 mg/g （1.25μmol/L atorvastatin） and 98.03 ± 10.24 mg/g （5μmol/L atorvastatin）. Conclusion Atorvastatin may downregulate ox-LDL-induced CD36 expression and decrease lipid accumulation in THP- 1 macrophages.