摘要
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。
Objective： To observe the effect of C-C motif ligand 2 （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. HUVECs were plated into 6-well plate, when cells were grown to 80-90 %, CCL2 overexpression plasmid [pcDNA3.1 （＋）-CCL2] or CCL2 small interfering RNA （si-CCL2） was transfected into HUVECs. At different time points after transfection （12 h, 24 h and 48 h）, HUVECs were collected for RNA and protein extraction. The mRNA expressions of CCL2 and ICAM-1 were examined using quantitative RT-PCR. The protein expressions of CCL2 and ICAM-1 were examined using western blotting. Results： （1） Compared with pcDNA3.1 （＋） group, the expressions of CCL2 mRNA and protein were significantly higher in pcDNA3.1 （＋）-CCL2 group; Compared with si-Control group, the expressions of CCL2 mRNA and protein were significantly decreased in si-CCL2 group. （2） Compared with pcDNA3.1 （＋） group, pcDNA3.1 （＋）-CCL2 significantly increased the mR_NA and protein expressions of ICAM-1 in ~3VECs, and si-CCL2 significantly inhibited the mRNA and protein expressions of ICAM-1 in HUVECs compared with si-Control group. Conclusions： CCL2 could affect ICAM-1 expression in HUVECs, which may provide a theoretical basis for clarifying the mechanisms of atherosclerosis.
出处
《现代生物医学进展》
CAS
2016年第26期5024-5027,5037,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金青年基金项目(81500282)
关键词
趋化因子配体2
人脐静脉内皮细胞
细胞间粘附分子-1
动脉粥样硬化
C-C motif ligand 2 （CCL2）
Human umbilical vein endothelial ceils （HUVECs）
Intercellular adhesion molecule-I （ICAM- 1）
Atherosclerosis
