不对称二甲基精氨酸对THP-1源性巨噬细胞表达MIF和分泌TNF-α、IL-8的影响(英文)

目的观察不对称二甲基精氨酸(ADMA)对THP-1源性巨噬细胞表达巨噬细胞移动抑制因子(MIF)和分泌TNF-α、IL-8的影响。方法用PMA诱导单核细胞THP-1分化为巨噬细胞。将分化后的THP-1源性巨噬细胞用不同浓度的ADMA作用24 h后,分别采用RT-PCR、Western blot和ELISA方法检测MIF mRNA和蛋白表达水平及培养上清中TNF-α与IL-8的含量。结果 ADMA剂量依赖性的上调了THP-1源性巨噬细胞内MIF mRNA和蛋白表达,15μmol/L ADMA对MIFmRNA和蛋白表达的上调作用最明显。随着ADMA浓度增加,THP-1源性巨噬细胞培养上清中TNF-α和IL-8的水平逐渐升高,在15μmol/LADMA作用时达到峰值。结论 ADMA能够上调巨噬细胞内MIF的表达,促进TNF-α和IL-8的分泌。

Effect of asymmetric dimethylarginine on MIF expression and TNF-α and IL-8 secretion in THP-1 monocytes-derived macrophages

Objective To investigate the effect of ADMA on macrophage migration inhibitory factor (MIF) expression and tumor necrosis factor-α (TNF-α) and IL-8 secretion in THP-Ⅰ monocyte-derived macrophages. Methods THP-1 monocytes were induced to differentiate into macrophages by a 24-h incubation with 160 nmol/L PMA. The THP-1 monocyte-derived macrophages were exposed to different concentrations of ADMA for 24 h, and the changes in MIF mRNA and protein expressions were analyzed with RT-PCR and Western blotting, respectively. Enzyme-linked immunosorbent assay was used to detect the levels of TNF-α and IL-8 in the supernatant of THP-l-derived macrophages following ADMA treatments. Results ADMA obviously up-regulated MIF mRNA and protein expressions in THP-1-derived macrophages in a concentrationdependent manner. Exposure of the cells to 15 μ mol/L ADMA for 24 h showed the most potent effect in up-regulating MIF mRNA and protein expressions. ADMA treatment also resulted in a dose-dependent increase of the levels of TNF-a and IL-8 in the culture supernatant of the macrophages, and the peak levels occurred following the treatment with 15 μ mol/L ADMA.Conclusion ADMA can up-regulate MIF expression and induce TNF-α and IL-8 secretion in THP-1 monocyte-derived macrophages.