趋化因子CCL2对人脐静脉内皮细胞ICAM-1 表达的影响

目的:探讨CC类趋化因子配体2(C-C motif ligand 2,CCL2)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)中细胞间粘附分子-1(intercellular adhesion molecule-1,ICAM-1)表达的影响。方法:体外分离培养HUVECs细胞,将HUVECs铺至6孔板中,待细胞融合至80-90%时,将CCL2过表达载体[pc DNA3.1(+)-CCL2]及CCL2小分子干扰RNA(si-RNA)分别转染到HUVECs中,于转染后12 h、24 h和48 h收集细胞进行RNA及蛋白提取。荧光定量PCR方法检测HUVECs中CCL2及ICAM-1基因m RNA表达。Western blotting检测HUVECs中CCL2及ICAM-1蛋白表达。结果:(1)与pc DNA3.1(+)组相比较,pc DNA3.1(+)-CCL2组中CCL2基因m RNA和蛋白水平均显著升高;与si-Control组相比较,si-CCL2组中CCL2基因m RNA和蛋白表达均明显下降。(2)与对照组比较,pc DNA3.1(+)-CCL2组明显增加HUVECs中ICAM-1的m RNA及蛋白表达,而si-CCL2组显著抑制HUVECs中ICAM-1的m RNA及蛋白表达。结论:CCL2能增加HUVECs中ICAM-1基因m RNA和蛋白表达,为深入认识动脉粥样硬化的发病机制提供了理论依据。

Effect of Chemokine CCL2 on the ICAM-1 Expression of HUVECs

Objective:To observe the effect of C-C motif ligand2 (CCL2) on the expression of intercellular adhesion molecule-1(ICAM-1) in human umbilical vein endothelial cells (HUVECs).Methods:HUVECs cells were isolated and cultured in vitro. HUVECswere plated into 6-well plate, when cells were grown to 80-90 %, CCL2 overexpression plasmid [pcDNA3.1 (+)-CCL2] or CCL2 smallinterfering RNA (si-CCL2) was transfected into HUVECs. At different time points after transfection (12 h, 24 h and 48 h), HUVECs werecollected for RNA and protein extraction. The mRNA expressions of CCL2 and ICAM-1 were examined using quantitative RT-PCR. Theprotein expressions of CCL2 and ICAM-1 were examined using western blotting.Results:(1) Compared with pcDNA3.1 (+) group, theexpressions of CCL2 mRNA and protein were significantly higher in pcDNA3.1 (+)-CCL2 group; Compared with si-Control group, theexpressions of CCL2 mRNA and protein were significantly decreased in si-CCL2 group. (2) Compared with pcDNA3.1 (+) group,pcDNA3.1 (+)-CCL2 significantly increased the mRNA and protein expressions of ICAM-1 in HUVECs, and si-CCL2 significantlyinhibited the mRNA and protein expressions of ICAM-1 in HUVECs compared with si-Control group.Conclusion:CCL2 could affectICAM-1 expression in HUVECs, which may provide a theoretical basis for clarifying the mechanisms of atherosclerosis.