Yasui Hirofumi, Andoh Akira, Bamba Shigeki, Inatomi Osamu, Ishida Hideaki, Fujiyama Yoshihide
Department of Internal Medicine, Shiga University of Medical Science, Seta-Tukinowa, Otsu, Japan.
Digestion. 2004;69(1):34-44. doi: 10.1159/000076545. Epub 2004 Jan 30.
The coordinated expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) plays a crucial role in tissue remodeling. We investigated the effects of fibroblast growth factor (FGF)-2 on the secretion of MMPs and TIMPs in human intestinal subepithelial myofibroblasts (SEMFs).
The secretion of MMP-s and TIMPs was determined by ELISA or Western blotting. The mRNA expression of MMPs and TIMPs was assessed by Northern blotting. The activating protein (AP)-1-DNA binding activity was evaluated by electrophoretic gel mobility shift assays (EMSA).
Unstimulated intestinal SEMFs constitutively secreted MMP-2 and TIMP-2. FGF-2 stimulated MMP-1, MMP-3 and TIMP-1 secretion, but did not affect MMP-2 or TIMP-2 secretion. FGF-2 induced AP-1-DNA binding activity, and the c-Jun/AP-1 inhibitor curcumin attenuated the FGF-2-induced MMP-1, -3 and TIMP-1 mRNA expression. Mitogen-activated protein (MAP) kinase inhibitors (U0126 and PD098059) also blocked the MMP-1, -3 and TIMP-1 secretion. Furthermore, FGF-2 dose-dependently induced FGF-2 mRNA expression in these cells.
FGF-2 may be one of important regulatory factors for extracellular matrix turnover via a modulation of MMP and TIMP secretion from SEMFs.
基质金属蛋白酶（MMPs）与金属蛋白酶组织抑制剂（TIMPs）的协同表达在组织重塑中起关键作用。我们研究了成纤维细胞生长因子（FGF）-2对人肠上皮下肌成纤维细胞（SEMFs）中MMPs和TIMPs分泌的影响。
通过酶联免疫吸附测定（ELISA）或蛋白质印迹法测定MMPs和TIMPs的分泌。通过Northern印迹法评估MMPs和TIMPs的mRNA表达。通过电泳凝胶迁移率变动分析（EMSA）评估激活蛋白（AP）-1与DNA的结合活性。
未受刺激的肠SEMFs组成性分泌MMP-2和TIMP-2。FGF-2刺激MMP-1、MMP-3和TIMP-1的分泌，但不影响MMP-2或TIMP-2的分泌。FGF-2诱导AP-1与DNA的结合活性，c-Jun/AP-1抑制剂姜黄素减弱FGF-2诱导的MMP-1、-3和TIMP-1 mRNA表达。丝裂原活化蛋白（MAP）激酶抑制剂（U0126和PD098059）也阻断MMP-1、-3和TIMP-1的分泌。此外，FGF-2剂量依赖性地诱导这些细胞中FGF-2 mRNA的表达。
FGF-2可能是通过调节SEMFs中MMP和TIMP的分泌来影响细胞外基质周转的重要调节因子之一。
