氧化型高密度脂蛋白对人单核细胞株THP-1细胞迁移及单核细胞趋化蛋白1含量的影响

Effects of Oxidized High Density Lipoprotein on Migration and Monocyte Chemoattractant Protein-1 of Human Monocytic THP-1 Cells

为研究氧化型高密度脂蛋白对单核细胞的趋化作用及单核细胞趋化蛋白 1含量的影响 ,进一步探讨氧化型高密度脂蛋白的致动脉粥样硬化机制 ,以一次性密度梯度超速离心法分离人血浆高密度脂蛋白 ,硫酸铜介导法进行氧化修饰 ,单核细胞为培养的人单核细胞株THP 1细胞 ,趋化实验采用改良Boyden趋化室 ,酶联免疫吸附检测法测定单核细胞趋化蛋白 1的含量。结果显示 ,氧化型高密度脂蛋白对THP 1细胞具有明显的趋化作用 ,其蛋白质浓度在 5 0和 10 0mg L时进入膜内的细胞数分别是对照组的 2倍 (P <0 .0 5 )和 3倍 (P <0 .0 1) ;雌二醇可部分抑制氧化型高密度脂蛋白对THP 1细胞的趋化作用。氧化型高密度脂蛋白可促进THP 1细胞单核细胞趋化蛋白 1的释放 ,氧化型高密度脂蛋白组单核细胞趋化蛋白 1的水平 ,约为对照组的 2倍 (P <0 .0 1) ;雌二醇可部分抑制氧化型高密度脂蛋白对THP 1细胞单核细胞趋化蛋白 1释放的促进作用。以上提示 ,氧化型高密度脂蛋白还可能通过促进循环血中单核细胞向血管内膜层迁移和聚集以及单核细胞趋化蛋白 1的释放而促进动脉粥样硬化的发生 。

Aim To study the effects of oxidized high density lipoprotein (ox HDL) on the migration of THP 1 cells and monocyte chemoattractant protein 1 (MCP 1)in the conditioned medium. Methods Human plasma lipoproteins were separated by one step ultracentrifugation with a simple density gradient, the in vitro oxidation of high density lipoprotein (HDL) was mediated by CuSO 4. THP 1 cells (a human acute monocytic leukemia cell line) were cultured for the experiments. The migration of THP 1 cells stimulated by oxidized high density lipoprotein was assayed with reformed Boyden. Results The migrated cells in the membrane (in Boyden well) were 52.5±6.5 in control group, 93.5±10.1 in 50 mg protein/L oxidized high density lipoprotein group(P<0.05), 148.5±21.6 in 100 mg protein/L oxidized high density lipoprotein group(P<0.01), 112.8±19.9 in oxidized high density lipoprotein+E 2 (17 β estradiol) group(P<0.05, compared with 100 mg protein/L oxidized high density lipoprotein group), and without significant changes in HDL groups. MCP 1 in the conditioned medium of THP 1 cells were as follows: 76.5±11.6 ng/g protein in control group, 141.0±19.5 ng/g protein in 100 mg protein/L oxidized high density lipoprotein group(P<0.01), 117.5±15.2 ng/g protein in oxidized high density lipoprotein+E 2 group(P<0.05 compared with oxidized high density lipoprotein group), and without significant change in HDL group. Conclusions oxidized high density lipoprotein could stimulate the migration of THP 1 cells and the production of MCP 1. It suggested that oxidized high density lipoprotein may participate in the primary pathogenesis of atherosclerosis by promoting the migration of monocyte to the intima of artery and increasing MCP 1; 17 β estradiol may have protective effect.