上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

GM-CSF通过JNK/c-Jun通路介导成纤维细胞MMP2/MMP9的表达

王宇星，严敏，彭银波，俞为荣，姚敏，方勇   

  1. 上海交通大学 医学院附属第三人民医院烧伤整形科， 上海 201999
  • 出版日期:2015-07-28 发布日期:2015-08-27
  • 通讯作者: 方勇, 电子信箱: fangyong1020@hotmail.com。
  • 作者简介:王宇星（1989—）， 女， 硕士生; 电子信箱: wangyx0424@outlook.com。
  • 基金资助:

    国家自然科学基金(81272081);上海市教育委员会科研创新项目（12ZZ112）

Induction of MMP2/MMP9 expression of fibroblasts by GM-CSF via JNK/c-Jun signaling pathway

WANG Yu-xing, YAN Min, PENG Yin-bo, YU Wei-rong, YAO Min,  FANG Yong   

  1. Department of Burn and Plastic Surgery, Shanghai Third People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China
  • Online:2015-07-28 Published:2015-08-27
  • Supported by:

    National Natural Science Foundation of China, 81272081; Innovation Program of Shanghai Municipal Education Commission, 12ZZ112

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摘要：

目的  探讨粒细胞单核巨噬细胞集落刺激因子（GM-CSF）对人皮肤成纤维细胞表达基质金属蛋白酶2和基质金属蛋白酶9（MMP2/MMP9）的影响及其相关机制。方法  ELISA法和明胶酶谱检测不同GM-CSF浓度（0、10、50、100和500 ng/mL）刺激下成纤维细胞上清液内MMP2/MMP9的浓度及活性；Western blotting和real-time PCR分别用于检测GM-CSF或SP600125+GM-CSF处理组成纤维细胞后MMP2/MMP9/JNK/p-JNK/c-Jun/p-c-Jun的蛋白水平以及MMP2/MMP9的mRNA水平；ELISA法检测GM-CSF或SP600125+GM-CSF处理成纤维细胞不同时间（6、12、24和48 h）后细胞上清内MMP2/MMP9的浓度。结果  GM-CSF可以显著诱导人皮肤成纤维细胞MMP2/MMP9的表达与分泌（P<0.05），且分泌的MMP2/MMP9浓度及活性与GM-CSF浓度呈正相关；GM-CSF处理成纤维细胞后，JNK及c-Jun蛋白的磷酸化水平和MMP2/MMP9的mRNA水平均高于对照组（P<0.05），若经SP600125预处理，GM-CSF诱导的c-Jun磷酸化水平则显著被抑制（P<0.05），且MMP2/MMP9的mRNA水平也明显降低（P<0.05）；GM-CSF诱导成纤维细胞MMP2/MMP9的表达在24 h时达到最高值，24～48 h逐渐降低（P<0.05）；若将SP600125与GM-CSF共处理成纤维细胞，MMP2的表达水平则显著低于对照组，MMP9的表达水平却略高于对照组。结论  GM-CSF能够通过活化JNK/c-Jun信号通路来诱导成纤维细胞MMP2/MMP9的表达和分泌。

关键词: 粒细胞单核巨噬细胞集落刺激因子, 基质金属蛋白酶2, 基质金属蛋白酶9, 人皮肤成纤维细胞, JNK/c-Jun通路

Abstract:

Objective  To investigate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the expressions of matrix metalloproteinases-2 and matrix metalloproteinases-9 (MMP2/MMP9) of human dermal fibroblasts and relevant mechanisms.  Methods  ELISA and Gelatin zymography were adopted to detect concentrations and activity of MMP2/MMP9 in the supernatant of fibroblasts treated by GM-CSF of different concentrations (0, 10, 50, 100, 500 ng/mL). The protein levels of MMP2, MMP9, JNK, p-JNK, c-Jun, and p-c-Jun and mRNA levels of MMP2 and MMP9 of fibroblasts treated by GM-CSF or SP600125+GM-CSF were detected by Western blotting and real-time PCR, respectively. The concentrations of MMP2/MMP9 in the supernatant of fibroblasts treated by GM-CSF or SP600125+GM-CSF at different time points (6, 12, 18, and 24 h) were detected by ELISA.  Results  GM-CSF could significantly induce expression and secretion of MMP2/MMP9 of human dermal fibroblasts (P<0.05) and the concentration and activity of secreted MMP2/MMP9 positively correlated with the concentration of GM-CSF. The phosphorylation of JNK and c-Jun and the mRNA level of MMP2/MMP9 of fibroblasts treated by GM-CSF were higher than those of controls (P<0.05). The phosphorylation of c-Jun and the mRNA level of MMP2/MMP9 induced by GM-CSF were significantly inhibited after being pretreated by SP600125 (P<0.05). The expression of MMP2/MMP9 of fibroblasts induced by GM-CSF reached the peak at 24 h and gradually decreased from 24 to 48 h (P<0.05). The expression of MMP2 of fibroblasts treated by both SP600125 and GM-CSF was significantly lower than that of controls, while the expression of MMP9 was slightly higher than that of controls.  Conclusion  GM-CSF can induced the expression and secretion of MMP2/MMP9 of fibroblasts by activating the JNK/c-Jun signaling pathway.

Key words: granulocyte-macrophage colony-stimulating factor, matrix metalloproteinases-2, matrix metalloproteinases-9, human dermal fibroblast, JNK/c-Jun signaling pathway