Xia Lin, Dong Zhaogang, Zhang Yun, Zhang Xiaoying, Song Xiaobin, Sun Mingxia, Hu Yingwei, Liu Shaohua, Wang Ketao, Qu Xun, Wei Fengcai
Department of Stomatology, Qilu Hospital, and Institute of Stomatology, Shandong University, Jinan, PR China; Department of Stomatology, Shuguang Branch of Shanghai Baoshan Hospital, Shanghai, PR China.
Institute of Basic Medical Sciences and Key Laboratory of Cardiovascular Proteomics in Shandong Province, Qilu Hospital, Shandong University, Jinan, PR China.
J Microbiol Immunol Infect. 2016 Jun;49(3):344-51. doi: 10.1016/j.jmii.2014.06.008. Epub 2014 Aug 12.
BACKGROUND/PURPOSE: Soluble vascular endothelial growth factor receptor-1 (sVEGFR1) antagonizes angiogenesis by inhibiting the biological function of vascular endothelial growth factor (VEGF). Immature dendritic cells (imDCs) express high levels of sVEGFR1 during development and are antiangiogenic. This study aimed to investigate the changes in VEGFR1, sVEGFR1, and VEGF levels during the development of imDCs and explore the underlying signaling mechanisms.
To model the differentiation of imDCs from monocytes, RAW264.7 cells, a murine monocyte/macrophage cell line, were stimulated by interleukin-4 (IL-4; 10 ng/mL, 20 ng/mL, and 40 ng/mL) and/or by granulocyte-macrophage colony-stimulating factor (GM-CSF; 10 ng/mL, 20 ng/mL, and 50 ng/mL) and/or pretreated by the p38 inhibitor SB203580. The levels of VEGFR1, sVEGFR1, and VEGF were detected by reverse transcription polymerase chain reaction (RT-PCR), Western blot, and enzyme-linked immunosorbent assay (ELISA).
IL-4 increased the VEGFR1 mRNA and sVEGFR1 levels in RAW264.7 (p < 0.05). This increase was inhibited by SB203580. Granulocyte-macrophage colony-stimulating factor increased the sVEGFR1 levels, but it had no significant effect on VEGFR1 mRNA levels. SB203580 decreased the expression of VEGFR1 mRNA induced by GM-CSF, whereas sVEGFR1 was unaffected. IL-4 had a greater effect on sVEGFR1 levels, compared to GM-CSF.
IL-4 and GM-CSF increased sVEGFR1 levels, but did not significantly effect VEGF expression, and led to the antiangiogenesis properties of monocytes. p38 Mitogen-activated protein kinase signaling has an important role in the process.
背景/目的:可溶性血管内皮生长因子受体-1(sVEGFR1)通过抑制血管内皮生长因子(VEGF)的生物学功能来拮抗血管生成。未成熟树突状细胞(imDCs)在发育过程中高水平表达sVEGFR1,具有抗血管生成作用。本研究旨在探讨imDCs发育过程中VEGFR1、sVEGFR1和VEGF水平的变化,并探索潜在的信号传导机制。
为模拟imDCs从单核细胞的分化过程,用白细胞介素-4(IL-4;10 ng/mL、20 ng/mL和40 ng/mL)和/或粒细胞-巨噬细胞集落刺激因子(GM-CSF;10 ng/mL、20 ng/mL和50 ng/mL)刺激小鼠单核细胞/巨噬细胞系RAW264.7细胞,和/或用p38抑制剂SB203580进行预处理。通过逆转录聚合酶链反应(RT-PCR)、蛋白质免疫印迹法和酶联免疫吸附测定(ELISA)检测VEGFR1、sVEGFR1和VEGF的水平。
IL-4增加RAW264.7细胞中VEGFR1 mRNA和sVEGFR1水平(p < 0.05)。这种增加被SB203580抑制。粒细胞-巨噬细胞集落刺激因子增加sVEGFR1水平,但对VEGFR1 mRNA水平无显著影响。SB203580降低GM-CSF诱导的VEGFR1 mRNA表达,而sVEGFR1不受影响。与GM-CSF相比,IL-4对sVEGFR1水平的影响更大。
IL-4和GM-CSF增加sVEGFR1水平,但对VEGF表达无显著影响,并导致单核细胞具有抗血管生成特性。p38丝裂原活化蛋白激酶信号传导在此过程中起重要作用。