非洛地平对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞黏附分子表达的影响

Effects of felodipine on the expression of cell adhesion molecule in human umbilical vein endothelial cells induced by oxidized low density lipoprotein

目的研究非洛地平对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞中细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)mRNA、蛋白表达影响及其抗动脉粥样硬化的作用机制。方法将氧化型低密度脂蛋白(ox-LDL)与人脐静脉内皮细胞(HUVECs)共孵育,筛选合适的ox-LDL干预浓度梯度与时间,与不同浓度的非洛地平(0.1,1,10μmol/L)共孵育24 h,real time-PCR检测细胞ICAM-1、VCAM-1 mRNA的表达,Western blot检测ICAM-1、VCAM-1蛋白表达变化。将HUVECs与U937单核细胞共培养,高倍镜下观察细胞形态变化,计数被黏附的HUVECs个数并计算黏附率。结果随着ox-LDL刺激浓度(6,12.5,25 mg/L)的增加,ICAM-1、VCAM-1 mRNA水平及蛋白表达逐渐上调,当ox-LDL浓度增加为25 mg/L时,作用最为显著(P<0.05)。非洛地平可抑制ICAM-1、VCAM-1 mRNA及蛋白水平的表达(P<0.05);25 mg/L ox-LDL处理时被黏附的HUVECs较空白对照明显增多(P<0.01);0.1μmol/L非洛地平显著抑制HUVECs与U937的相互黏附(P<0.01)。结论非洛地平通过下调人脐静脉内皮细胞ICAM-1、VCAM-1的表达从而抑制单核-血管内皮细胞黏附。

Objective To investigate the effect of felodipine on the mRNA and protein expression of intercellular adhesion molecule-1（ICAM-1）,vascular cell adhesion molecule-1（VCAM-1） in human umbilical vein endothelial cells induced by oxidized low density lipoprotein and its mechanism of anti-atherosclerosis.Methods The concentration gradient and time of incubation of oxidized low density lipoprotein（ ox-LDL） were screened,and then the human umbilical vein endothelial cells（ HUVECs） were co-incubated with ox-LDL and different concentration of felodipine（0.1,1,10 mol/L） for 24 h.Then the mRNA and protein expression levels of ICAM-1 and VCAM-1 were detected by real time-PCR and Western blot.After HUVECs and U937 cells were co-cultured,the morphological changes were observed under light microscope,and the number of HUVECs adhesion was counted and the adhesion rate was calculated.Results With the increase of ox-LDL concentration,the mRNA and protein expression levels of VCAM-1 and ICAM-1 gradually increased,and peaked at the concentration of 25 mg/L（ P0.05）.Felodipine suppressed the mRNA and protein expression of ICAM-1 and VCAM-1（ P0.05）.Adhesion rate of U937 cells to HUVECs were significantly higher in in 25 mg/L ox-LDL group than in 0mg/L ox-LDL group（ P0.01）.Felodipine（0.1 mol/L） significantly inhibited the adhesion of HUVECs and U937（ P0.01）.Conclusion felodipine may inhibit the adhesion of monocyte and endothelial cell through down-regulation of ICAM-1,VCAM-1 expression in endothelial cells and play an anti-inflammatory protective effect.