8-Br-cAMP对THP1细胞中ABCA1及单核细胞趋化因子的影响

 目的 以THP1细胞株为靶点,研究8-Br-cAMP对ABCA1、MCP-1基因mRNA和蛋白质表达及IL-1β蛋白质的影响,阐明ABCA1基因在AS及泡沫细胞形成中新的可能机制.方法 复苏培养THP1细胞株,加入8-Br-cAMP(0.5 mmol/L)刺激3、6、12、24 h,荧光定量RT-PCR和Western蛋白印迹法及ELISA法检测ABCA1、MCP-1及IL-1βmRNA和蛋白表达量;用ABCA1的反义寡核苷酸(100 nmol/L)转染THP1细胞,给予上述8-Br-cAMP的刺激,同样的方法观察上述指标的改变.结果 予8-Br-cAMP刺激后,THP1细胞中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后,8-Br-cAMP刺激后3、6 h ABCA1、MCP-1mRNA的表达降低,12、24 h ABCA1、MCP-1及IL-1β蛋白质的表达水平降低.结论 在8-Br-cAMP刺激下,THP1细胞中ABCA1可增加炎症因子表达,参与AS的发生.

Effect of 8-Br-cAMP on ABCA1 and MCP-1 in THP-1 cells

Objective To study the changes of ABCA1, MCP - 1 and IL - 1β after the treatment of THP1 cell lines with 8 - Br - cAMP, and to elucidate a new possible mechanism of ABCA1 gene in the formation of AS and foam cells. Method THP1 cells were resuscitated and cultured. 8 - Br - cAMP（0. 5mM） was added into the culture media and THP1 cells were harvested at indicated times. The mRNA and protein levels of ABCA1, MCP - 1 and IL - 115 were investigated by real - time quantitative RT - PCR, Western blotting and ELISA. Phosphorothioate oligonucleotides of ABCA1 were used at a final concentration of 100nmol/L for all the antisense transfection experiments. The same experiments were repeated after the transfection of oligonucleotides of ABCAI. Results The mRNA and protein expressions were increased after incubation with Ox - LDL. After transfection with antisense oligonucleotides of ABCA1, the mRNA decreased at 3 and 6 h, and the proteins decreased at 12 and 24 h. Conclusion ABCA1 increases expressions of inflammatory cytokines stimulated by 8 - Br - cAMP in THP - 1 cells and participates in atheroselerogenesis.