ERK通路在单核巨噬细胞集落刺激因子调控成纤维细胞血管内皮生长因子表达中的作用

目的 研究单核巨噬细胞集落刺激因子（GMCSF）对成纤维细胞血管内皮生长因子（VEGF）表达的影响，初步探讨细胞外信号调节激酶（ERK）通路对此过程的调控作用。方法 在对数生长期人皮肤成纤维细胞培养液中添加GMCSF和（或）ERK通路特异性抑制剂（PD98059）进行干预，实验分为空白对照组、PD98059组、GMCSF组及PD98059+GMCSF组。收集各组细胞及培养上清液，采用RT-PCR技术和Western blotting方法分别检测各组人皮肤成纤维细胞内VEGF mRNA和蛋白表达；ELISA法检测细胞培养上清液中VEGF蛋白分泌水平。结果 与空白对照组和PD98059组比较，GMCSF组人皮肤成纤维细胞内VEGF mRNA和蛋白相对表达量以及细胞培养上清液中VEGF蛋白质量体积浓度明显升高（P<0.05）；而在GMCSF+PD98059组中，人皮肤成纤维细胞内和细胞培养上清液中相应指标表达水平均显著低于GMCSF组（P<0.05）。结论 GMCSF可促进人皮肤成纤维细胞VEGF表达，ERK通路在此过程中发挥重要作用。

Role of ERK pathway in regulation of expression of vascular endothelial growth factor in fibroblasts by mononuclear macrophage colony-stimulating factor

Objective To investigate the effect of mononuclear macrophage colony-stimulating factor (GMCSF) on the expression of vascular endothelial growth factor (VEGF) in fibroblasts, and explore the role of extracellular signal-regulated kinase (ERK) pathway in this process. Methods GMCSF and/or ERK pathway specific inhibitor (PD98059) were added to the culture fluid of human skin fibroblasts at exponential phase of growth, and blank control group, PD98059 group, GMCSF group and PD98059+GMCSF group were divided. Cells and supernatant were harvested from each group, the expression of VEGF mRNA and protein in human skin fibroblasts of each group were detected by RT-PCR and Western blotting, respectively, and the levels of VEGF protein in the supernatant were determined by ELISA. Results Compared with blank control group and PD98059 group, the expression of VEGF mRNA and protein in human skin fibroblasts and the levels of VEGF protein in the supernatant in GMCSF group were significantly increased (P<0.05), while the parameters in GMCSF+PD98059 group were significantly lower than those in GMCSF group (P<0.05). Conclusion GMCSF can up-regulate the expression of VEGF in human skin fibroblasts, and ERK pathway plays an important role in this process.