Ox-LDL对主动脉平滑肌细胞中ABCA1及炎性因子表达的影响

 目的 研究氧化低密度脂蛋白(Ox-LDL)诱导作用下,ATP结合盒转运子A1(ABCA1)对人主动脉平滑肌中单核细胞趋化因子-1(MCP-1)、细胞间黏附分子-1(ICAM-1)mRNA和蛋白质及白介素-1β(IL-1β)表达的影响.方法 复苏培养人主动脉平滑肌细胞CC2571、Ox-LDL (30 μg/ml) 刺激3、6、12、24 h后,以荧光定量RT-PCR法检测ABCA1、ICAM-1、MCP-1mRNA表达,Western蛋白印迹法及ELISA法检测ABCA1、ICAM-1、MCP-1及IL-1β蛋白质表达;用ABCA1的反义寡核苷酸抑制ABCA1表达后,观察上述指标的变化.结果 予Ox-LDL刺激后, CC2571细胞中ABCA1、MCP-1、ICAM-1的mRNA和蛋白及IL-1β蛋白质表达均增加,反义寡核苷酸转染后3、6 h时上述指标的mRNA表达降低,12、24 h蛋白质表达降低.结论 Ox-LDL诱导主动脉平滑肌细胞炎性因子的表达,ABCA1可能通过增强上述炎性因子表达参与动脉粥样硬化(atherosclerosis,AS)的发生.

Effects of ABCA1 on human aortic smooth muscle cells' inflammatory cytokines induced by oxidized low density lipoprotein

Objective To evaluate the effect of ATP binding cassette transporter A1 （ ABCA1 ） on mRNA and on protein expression of intercellular adhesion molecule - 1 （ ICAM - 1 ） and monocyte chemoattractant protein - 1 （ MCP - 1 ） and protein expression of interleukin - 1β （ IL - 1 β ） in aortic smooth muscle cell CC2571 induced by oxidized low density lipoprotein （ Ox - LDL） in order to investigate the mechanism by which ABCA1 contributes to atherosclerogenesis. Methods Human aortic smooth muscle cell CC2571 was cultured , Ox - LDL （30g/ml） was added into culture media and cells were harvested at 3, 6, 12 and 24 hours, respec- tively. The mRNA of ABCA1, MCP - 1, ICAM - 1 and IL - 1β was investigated by RT - PCR, Western blotting and ELISA. The pro- tein of ABCA1, MCP - 1, ZCAM - 1 and ZC - 1β was investigated. After phosphorothioate oligonucleotides of ABCA1 mixture were added to culture media at a final concentration of 100nmol/L, the same experiment was repeated. Results The mRNA and protein a- mounts of ABCA1, MCP- 1 and ICAM -1 and also protein amount of IL - 1 β were increased after vascular smooth muscle cells were incubated with Ox - LDL. the expression of mRNA, when transfected with antisense oligonucleotides of ABCA1 was decreased at 3 and 6 hours, and protein expression decreased at 12 and 24 hours. Conclusions ABCA1 contributes to atherosclerogenesis by increasing the expression of inflammatory cytokines induced by Ox - LDL in hunman vascular smooth muscle cells.