氨氯地平对平滑肌细胞泡沫化过程中亲环素A表达和胆固醇蓄积的影响

目的 观察氨氯地平预处理对氧化型低密度脂蛋白诱导的平滑肌细胞泡沫化过程中亲环素A表达的影响及其对胆固醇蓄积的作用,从新的角度探讨氨氯地平的抗动脉粥样硬化作用.方法 实验分为6组:对照组、氧化型低密度脂蛋白组、氨氯地平(0.1、1.0和10.0 μmol/L)处理细胞1 h后加入80 mg/L氧化型低密度脂蛋白共同孵育72 h组及10.0 μmol/L氨氯地平单独处理组,Western-blot和RT-PCR分别检测亲环素A mRNA和蛋白质的表达改变;油红O染色观察细胞内脂滴的形成情况,高效液相色谱法检测细胞内胆固醇含量变化.结果 氧化型低密度脂蛋白处理组的细胞内亲环素A的表达明显减少;经氨氯地平处理后,随着氨氯地平浓度的增加,细胞内亲环素A的表达逐渐增加,10.0μmoL/L氨氯地平预处理细胞组效果最明显,较氧化型低密度脂蛋白组差异有显著性(P<0.05).油红O染色显示,氧化型低密度脂蛋白组细胞内大量脂滴形成,细胞内胆固醇酯/总胆固醇的比值为57.9%,符合泡沫细胞特征;预先予氨氯地平处理后,随着药物浓度的增加,细胞内脂滴逐渐减少,细胞内胆固醇酯/总胆固醇的比值逐渐降低,10.0μmol/L氨氯地平预处理细胞组效果最明显,为37.8%;结论 氨氯地平预处理,可上调氧化型低密度脂蛋白诱导的平滑肌细胞泡沫化过程中亲环素A的表达,减轻细胞内的胆固醇蓄积.

Effect of Amlodipine on Cyclophilin A Expression and Cellular Cholesterol Accumulation in Foam Cells Derived from Smooth Muscle Cell

Aim To study the function of amlodipine on cellular cholesterol accumulation and cyclophilin a ex-pression in foam cells derived from smooth muscle cell induced by oxidized low density lipoprotein (ox-LDL) to explore the new antiatherosclerotic role of Amlodipine. Methods The experiment groups included 6 groups:control group, ox-LDL group, different concentration(0.1, 1.0, 10.0μmoL/L)of amlodipine pretreated for 1 h and treated with 80 mg/L ox-LDL for 72 h group and amlodipine treated alone without ox-LDL group. RT-PCR and Western-blot analysis were used to detect the mRNA and protein expression of cyclophilin A. Oil red O dye experiment showed the lipid droplets in the cells of each group. High Performance liquid chromatography (HPLC) analysis was peformed to measure the content of cellular total cholesterol and cholesterol ester. Results RT-PCR and Western-blot analysis showed that with ox-LDL and am-lodipine treatment, cyclophilin A expression were greatly increased in a concentration-dependent manner. Oil Red O dye showed that with the treatment of amlodipine, the lipid droplets were greatly decreased in a concentration-dependent manner vs ox-LDL group. The content of cellular total cholesterol and cholesterol ester were both significantly increased with ox-LDL treatment but decreased after amlodipine treatment. Conclusion Amlodipine can up-regulate the expression of cyclophilin A and relieve the cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL.