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

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

作　　者：王宇星[1] 严敏[1] 彭银波[1] 俞为荣[1] 姚敏[1] 方勇[1] 

机构地区：[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　　处：《上海交通大学学报（医学版）》2015年第7期933-939,共7页Journal of Shanghai Jiao tong University:Medical Science

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

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/m L）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。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 M MP2/M MP9 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,and24 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.

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 

分 类 号：R751[医药卫生—皮肤病学与性病学]