摘要
目的研究在致动脉粥样硬化(AS)因子氧化型低密度脂蛋白(Ox-LDL)刺激下,人血管内皮细胞ECV304中腺苷三磷酸结合盒转运体A1(ABCA1)基因对炎性因子单核细胞趋化蛋白1(MCP-1)的调节作用,从而阐明ABCA1基因在AS发生中的作用机制。方法培养人血管内皮细胞ECV304,加入Ox-LDL(30 ng/ml)刺激3、6、12、24 h,以荧光定量RT-PCR检测ABCA1、MCP-1 mRNA表达量,Western蛋白印迹法和ELISA法检测ABCA1、MCP-1及IL-1β蛋白表达量;ABCA1的反义寡核苷酸(100 nmol/L)转染人血管内皮细胞,给予上述的Ox-LDL,同样方法测定上述指标的改变。结果人血管内皮细胞ECV304在给予Ox-LDL刺激后,ABCA1、MCP-1的mRNA和蛋白质水平及IL-1β蛋白质水平均增高;给予反义寡核苷酸转染3、6 h后,ABCA1、MCP-1 mRNA的表达降低,12、24h后ABCA1、MCP-1及IL-1β蛋白质的表达水平降低。结论在Ox-LDL作用下,血管内皮细胞中的ABCA1可能通过增加IL-1β的表达,促进MCP-1释放,从而在AS的早期发生中发挥作用。
Objective To investigate the effects of ABCA1 on MCP-1 in vascular endothelial cell ECV304 after the treatment with Ox-LDL in order to find the new possible mechanisms that ABCA1 contributed to atherosclerogenesis.Methods Ox-LDL（30 ng/ml） was added into culture media and ECV304 cells were harvested at 3,6,12 and 24 h.The mRNA and protein levels of ABCA1,MCP-1 and IL-1β were investigated by real-time fluorescent quantitative RT-PCR,Western blot and ELISA methods.After phosphorothioate antisense oligonucleotides of ABCA1 mixtures were added to culture media at a final concentration of 100 nmol/L,the same experiments were repeated.Results The mRNA and protein of ABCA1,MCP-1 and IL-1β increased after the incubated with Ox-LDL.After transfection with antisense oligonucleotides of ABCA1,the mRNA of ABCA1 and MCP-1 decreased after 3 and 6 hours,and protein of ABCA1,MCP-1 and IL-1β deereased after 12 or 24 hours.Conclusion ABCA1 could probabley increase expressions of MCP-1 through IL-1β in human vascular endothelial cell after Ox-LDL stimulation and take effects on atherosclerogenesis.
出处
《山东医药》
CAS
北大核心
2010年第17期16-18,共3页
Shandong Medical Journal
基金
国家自然科学基金(30171028)
广东省自然科学基金(010616)
