巨噬细胞中ABCA1对炎症因子的调节及其意义

目的研究在8-溴-环磷酸腺苷（8-Br-cAMP）刺激下，巨噬细胞中三磷酸腺苷结合盒转运子（ABCA1）对细胞间黏附分子-l（ICAM-1）、单核细胞化学趋向蛋白-1（MCP-1）及白介素-1β（IL-1β）的调节作用，在细胞水平证实ABCAl在动脉粥样硬化发生巾的作用机制？方法用氟波酯（PMA）刺激THP-1细胞使之转变为巨噬细胞，8-Br-cAMP（0．5mmol／L）刺激3、6、12、24h后，以荧光定量RT-PCR和Western蛋白印迹法及ELISA法检测ABCAl、ICAM-1、MCP-1及IL-1BmRNA和蛋白质表达量；用反义寡核苷酸（100nmol／L）抑制ABCAl的表达，观察Ox-LDL刺激下上述指标的改变。结果予8-Br-cAMP刺激6、12h后，巨噬细胞ABCAl、ICAM-1、MCP-1mRNA和蛋白质水平及IL-1B蛋白质水平均增高：反义寡核甘酸转染巨噬细胞，8．Br—cAMP刺激后3、6h，ABCA1、ICAM-1、MCP．1mRNA的表达降低，刺激后12，24h，ABCA1、ICAM．1、MCP-1及IL-1B蛋白质的表达水平降低。结论在8-Br-cAMP刺激下，巨噬细胞ABCA1可增加炎症因子表达，参与动脉粥样硬化的发生。

Modulation of inflammatory cytokines by ATP-binding cassette A1 in THP-1 macrophages

OBJECTIVE: To observe the effects of ATP-binding cassette A1 (ABCA1) on intercellular cell adhesion molecule type 1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1) and interleukin-1beta (IL-1beta) in THP-1 macrophages stimulated with 8-Br-cAMP to identify a possible new mechanism that ABCA1 contributes to atherosclerogenesis (AS). METHODS: Monocytic THP-1 cells were cultured in the presence of 100 nmol/L phorbol myristate acetate (PMA) for 72 h to transform the cells into THP-1 macrophages. After the macrophages were stimulated with 8-Br-cAMP (final concentration 0.5 mmol/L) for 3, 6, 12 and 24 h respectively, the amounts of ABCA1, ICAM-1 and MCP-1 mRNA were examined by real-time fluorescent quantitative RT-PCR, and the protein amounts of ABCA1, ICAM-1, MCP-1 and IL-1beta were determined by Western blotting and enzyme-linked immunosorbent assay (ELISA). Phosphorothioate antisense oligonucleotides of ABCA1 were add into the culture media at a final concentration of 100 nmol/L and the experiments were repeated. RESULTS: ABCA1, ICAM-1 and MCP-1 mRNA and protein and IL-1beta protein were increased in the macrophages after stimulation with 8-Br-cAMP for 6 and 12 h. The mRNA expressions of ABCA1, ICAM-1 and MCP-1 were decreased significantly at 3 and 6 h (P<0.01), and the protein expressions of ABCA1, ICAM-1, MCP-1 and IL-1beta declined significantly at 12 and 24 h (P<0.01) after transfection of the macrophages with antisense oligonucleotides of ABCA1. CONCLUSION: ABCA1 can increase the expressions of the inflammatory cytokines in THP-1 macrophages stimulated by 8-Br-cAMP and plays a role in the pathogenesis of AS.