巨噬细胞ABCA1对Ox-LDL诱导的炎症因子 的调节及其意义.DOCVIP

临床医学论文-巨噬细胞ABCA1对Ox-LDL诱导的炎症因子 的调节及其意义 ???????????? 作者：郭志刚， 吴平生， 李建华， 赖文岩 【摘要】? 　　【目的】 研究在氧化低密度脂蛋白（Ox-LDL）诱导作用下，ATP结合盒转运子A1（ABCA1）对巨噬细胞中细胞间黏附分子－1（ICAM-1）、单核细胞化学趋向蛋白－1（MCP-1）mRNA和蛋白质及白介素－1β（IL-1β）蛋白质表达的影响，在细胞水平证实ABCA1对动脉粥样硬化的影响。【方法】 用氟波酯(PMA)刺激THP-1细胞使之转变为巨噬细胞，Ox-LDL (30 ?滋g/mL)刺激3、6、12、24 h后，以荧光定量RT－PCR和Western蛋白印迹法及酶联免疫吸附法（ELISA）检测ABCA1、ICAM-1、MCP-1及IL-1β mRNA和蛋白质表达量；用反义寡核苷酸（100 nmol/L）抑制ABCA1的表达，观察Ox-LDL刺激下上述指标的改变。【结果】 给予Ox-LDL刺激后，巨噬细胞ABCA1、ICAM-1、MCP-1的mRNA和蛋白及IL-1β蛋白质表达均增高；给予反义寡核苷酸转染后Ox-LDL刺激3、6 h，上述指标的mRNA表达降低(P0.01)，12、24 h蛋白质表达降低(P0.01)。【结论】 在巨噬细胞，ABCA1可增加Ox-LDL诱导的炎症因子表达，参与动脉粥样硬化的发生。 【关键词】? ATP结合盒转运子A1； 氧化低密度脂蛋白； 巨噬细胞； 动脉粥样硬化； 炎症因子 ??? Abstract:【Objective】 To evaluate the effects of ATP binding cassette transporter A1? (ABCA1) on mRNA and protein expression of intercellular adhesion molecule-1? (ICAM-1) and monocyte chemoattractant protein-1? (MCP-1) and protein expression of interleukin-1β (IL-1β) in THP-1 macrophage induced by oxidized low density lipoprotein? (Ox-LDL) in order to investigate? the mechanism? that ABCA1? contributes to atherosclerogenesis.【Methods】 Monocytic THP-1 cells were cultured with 100? nmol/L phorbol myristate acetate (PMA) for 72 hours to lead cells into THP-1 macrophage. Ox-LDL (30 ?滋g/mL) was added into culture media and THP-1 macrophage cells were harvested at 3, 6, 12, and 24 hours, respectively. The mRNA and protein levels of ABCA1, ICAM-1, MCP-1 and IL-1β were investigated by real-time fluorescent quantitative RT-PCR, Western blot, and ELISA methods. After phosphorothioate oligonucleotides of ABCA1 mixture were add to culture media at final concentration of 100? nmol/L, the same experiments were repeated. 【Results】 The mRNA and protein amounts of ABCA1, ICAM-1, and MCP-1 and also protein amount of IL-1β were increased after macrophage incubated with Ox-LDL. Transfected with antisense oligonucleotides of ABCA1, the expressive levels of mRNA were decreased at 3 and 6 hours (P 0.01), and protein at 12 and 24 hours (P 0.01). 【Con