Cyclooxygenase-2-regulated vascular endothelial growth factor release in gastric fibroblasts
Cyclooxygenase-2-regulated vascular endothelial growth factor release in gastric fibroblasts
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DOI：
10.1152/ajpgi.00537.2003
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发表时间：
2004-08-01
影响因子：
4.5
通讯作者：
Sakamoto, C
中科院分区：
文献类型：
--
作者：
Miura, S;Tatsuguchi, A;Sakamoto, C
VEGF是一种高度特异性的内皮细胞刺激因子，在组织再生过程中可能在血管生成中发挥重要作用。我们之前的研究表明，在溃疡床的间充质细胞中表达的环氧合酶-2 （COX-2）参与溃疡修复过程。为了明确COX-2在胃溃疡愈合过程中血管生成中的作用，我们在体内和体外研究了人胃成纤维细胞COX-2表达与VEGF生成的关系。胃成纤维细胞在RPMI 1640中培养，有或没有il -1 α或il -1 β，存在或不存在NS-398（一种选择性COX-2抑制剂）。酶联免疫吸附法检测上清VEGF和PGE(2)浓度。Western blot检测成纤维细胞中COX-2的表达。用免疫组织化学方法检测人胃溃疡手术切除组织中VEGF和COX-2的表达。IL-1剂量依赖性地促进培养胃成纤维细胞在刺激24小时后的VEGF释放。IL-1也通过COX-2诱导刺激胃成纤维细胞产生PGE(2)。NS-398显著抑制il -1刺激的胃成纤维细胞中VEGF和PGE(2)的释放；同时添加PGE(2)恢复ns -398抑制的VEGF释放。COX-2和VEGF免疫反应性共定位于胃组织溃疡床的成纤维细胞样细胞。这些结果提示，在IL-1体外刺激下，COX-2在胃成纤维细胞的VEGF生成中起关键作用，COX-2-VEGF途径诱导的血管生成可能参与胃溃疡愈合。
VEGF is a highly specific stimulator of endothelial cells and may play an important role in angiogenesis in the process of tissue regeneration. We previously showed that cyclooxygenase-2 (COX-2) expressed in mesenchymal cells of the ulcer bed is involved in the ulcer repair process. To clarify the role of COX-2 in angiogenesis during gastric ulcer healing, we investigated the relation between COX-2 expression and VEGF production in human gastric fibroblasts in vivo and in vitro. Gastric fibroblasts were cultured in RPMI 1640 with and without IL-1alpha or IL-1beta in the presence or absence of NS-398, a selective COX-2 inhibitor. Supernatant VEGF and PGE(2) concentrations were measured by enzyme-linked immunosorbent assay. COX-2 expression in fibroblasts was determined by Western blot analysis. VEGF and COX-2 expression in surgical resections of human gastric ulcer tissue was examined immunohistochemically. IL-1 dose dependently enhanced VEGF release in cultured gastric fibroblasts after a 24-h stimulation. IL-1 also stimulated PGE(2) production in gastric fibroblasts via COX-2 induction. NS-398 significantly suppressed VEGF and PGE(2) release from IL-1-stimulated gastric fibroblasts; concurrent addition of PGE(2) restored NS-398-inhibited VEGF release. COX-2 and VEGF immunoreactivity were colocalized in fibroblast-like cells in the ulcer bed of gastric tissues. These results suggest that COX-2 plays a key role in VEGF production in gastric fibroblasts stimulated by IL-1 in vitro and that angiogenesis induced by the COX-2-VEGF pathway might be involved in gastric ulcer healing.