Exendin-4 ameliorates oxidized-LDL-induced inhibition of macrophage migration in vitro via the NF-κB pathway
Exendin-4 ameliorates oxidized-LDL-induced inhibition of macrophage migration in vitro via the NF-κB pathway
复制标题
Exendin-4 通过 NF-κB 途径改善氧化 LDL 诱导的体外巨噬细胞迁移抑制
DOI：
10.1038/aps.2013.128
复制
发表时间：
2014-02-01
影响因子：
8.2
通讯作者：
Yao, Wen-bing
中科院分区：
文献类型：
--
作者：
Ma, Ge-fei;Chen, Song;Yao, Wen-bing
目的：探讨胰高血糖素样肽-1 （GLP-1）受体激动剂exendin-4对氧化低密度脂蛋白（ox-LDL）诱导的巨噬细胞迁移抑制的影响及其作用机制。方法：用3%巯基乙酸酯（2ml, ip）处理小鼠腹腔，提取原代腹腔巨噬细胞。用细胞迁移法检测巨噬细胞的迁移。采用半定量RT-PCR检测巨噬细胞迁移相关因子，包括瘦素样ox-LDL受体（LOX-1）、环氧化酶2 （COX-2）、肿瘤坏死因子(TNF)-α、白细胞介素-1 (IL-1) β、基质金属蛋白酶-2 （MMP-2）、细胞间粘附分子(ICAM)-1和巨噬细胞迁移抑制因子（MIF）。ELISA法检测MIF和ICAM-1蛋白的表达。明胶酶谱法测定MMP-9活性。通过共聚焦激光扫描显微镜观察NF-κB通路的激活情况。结果：ox-LDL （50 μg/mL）处理巨噬细胞可明显抑制巨噬细胞的迁移。此外，ox-LDL处理显著增加巨噬细胞迁移相关因子的表达、MMP-9的活性和NF-κB p65亚基的易位。特异性NF-κB抑制剂吡咯烷二硫代氨基甲酸铵（100 μmol/L）预处理可显著改善ox-LDL的上述作用。exendin-4预处理（25和50 nmol/L）也能显著改善ox-LDL的这些作用。结论：Exendin-4通过抑制ox-LDL诱导的ICAM-1和MIF的表达，改善ox-LDL对巨噬细胞迁移的体外抑制作用，这可能与NF-κB通路有关。
Aim:To investigate the effects of the glucagon-like peptide-1 (GLP-1) receptor agonist exendin-4 on oxidized low-density lipoprotein (ox-LDL)-induced inhibition of macrophage migration and the mechanisms underlying the effects of exendin-4.Methods:Primary peritoneal macrophages were extracted from the peritoneal cavity of mice treated with 3% thioglycollate (2 mL, ip). Migration of the macrophages was examined using a cell migration assay. Macrophage migration-related factors including leptin-like ox-LDL receptor (LOX-1), cyclooxygenase 2 (COX-2), tumor necrosis factor (TNF)-α, interleukin-1 (IL-1) β, matrix metalloproteinase-2 (MMP-2), intercellular adhesion molecule (ICAM)-1 and macrophage migration inhibitory factor (MIF) were measured using semi-quantitative RT-PCR. Expression of MIF and ICAM-1 proteins was examined with ELISA. Gelatin zymography was used to evaluate the activity of MMP-9. Activation of the NF-κB pathway was determined by confocal laser scanning microscopy.Results:Treatment of the macrophages with ox-LDL (50 μg/mL) markedly suppressed the macrophage migration. Furthermore, ox-LDL treatment substantially increased the expression of the macrophage migration-related factors, the activity of MMP-9 and the translocation of the NF-κB p65 subunit. These effects of ox-LDL were significantly ameliorated by pretreatment with the specific NF-κB inhibitor ammonium pyrrolidine dithiocarbamate (100 μmol/L). These effects of ox-LDL were also significantly ameliorated by pretreatment with exendin-4 (25 and 50 nmol/L).Conclusion:Exendin-4 ameliorates the inhibition of ox-LDL on macrophage migration in vitro, via suppressing ox-LDL-induced expression of ICAM-1 and MIF, which is probably mediated by the NF-κB pathway.