Szabó I L, Pai R, Soreghan B, Jones M K, Baatar D, Kawanaka H, Tarnawski A S
Gastroenterology Section (111G), DVA Medical Center Long Beach (CA), 5901 East Seventh Street, Long Beach, CA 90822, USA.
J Physiol Paris. 2001 Jan-Dec;95(1-6):379-83. doi: 10.1016/s0928-4257(01)00051-1.
Nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, indomethacin (IND), ibuprofen and newer cyclooxygenase-2 selective NSAIDs (e.g. celecoxib) delay gastric ulcer healing partly through the inhibition of angiogenesis, but the molecular mechanisms involved are not fully elucidated. Effective angiogenesis is required for ulcer healing to supply oxygen and nutrients to the healing site. The early growth response factor (Egr-1) is a transcription factor, which is rapidly activated by a variety of extracellular signals or tissue injury and is important for angiogenesis to occur. This study aimed to determine whether indomethacin (IND) and/or the selective COX-2 inhibitor, NS-398, interfere with egr-1 gene expression in human microvascular endothelial cells (HMVEC) in response to vascular endothelial growth factor (VEGF) stimulation. HMVEC were treated with 0.5 mM IND or 100 microM NS-398 for 16 h, and then VEGF (10 ng/ml) or vehicle was added. Egr-1 mRNA and protein expression levels were determined by RT-PCR and Western-blotting, respectively. VEGF treatment caused a significant elevation of Egr-1 mRNA (261+/-21%, P<0.001) and protein expression (174+/-15%, P<0.01) vs. vehicle. IND pre-treatment significantly inhibited VEGF-induced Egr-1 mRNA expression by 29+/-4% (P<0.01) and protein expression by 41+/-8% (P<0.05). NS-398 inhibited VEGF-induced Egr-1 mRNA and protein expression by 23+/-3% and 35+/-4%, respectively (both P<0.01). Since transcriptional activation of egr-1 is responsible for expression of proteins involved in proliferation of endothelial cells essential for angiogenesis, these results provide a new mechanism for NSAIDs' interference with angiogenesis.
非甾体抗炎药（NSAIDs），如阿司匹林、吲哚美辛（IND）、布洛芬以及新型环氧化酶-2选择性NSAIDs（如塞来昔布），部分通过抑制血管生成来延缓胃溃疡愈合，但其涉及的分子机制尚未完全阐明。有效的血管生成是溃疡愈合所必需的，可为愈合部位提供氧气和营养物质。早期生长反应因子（Egr-1）是一种转录因子，可被多种细胞外信号或组织损伤迅速激活，对血管生成的发生至关重要。本研究旨在确定吲哚美辛（IND）和/或选择性COX-2抑制剂NS-398是否会干扰人微血管内皮细胞（HMVEC）中egr-1基因的表达，以应对血管内皮生长因子（VEGF）的刺激。将HMVEC用0.5 mM IND或100 microM NS-398处理16小时，然后加入VEGF（10 ng/ml）或溶剂。分别通过逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法测定Egr-1 mRNA和蛋白质表达水平。与溶剂相比，VEGF处理导致Egr-1 mRNA（261±21%，P<0.001）和蛋白质表达（174±15%，P<0.01）显著升高。IND预处理显著抑制VEGF诱导的Egr-1 mRNA表达29±4%（P<0.01）和蛋白质表达41±8%（P<0.05）。NS-398分别抑制VEGF诱导的Egr-1 mRNA和蛋白质表达23±3%和35±4%（均P<0.01）。由于egr-1的转录激活负责血管生成所必需的内皮细胞增殖相关蛋白质的表达，这些结果为NSAIDs干扰血管生成提供了一种新机制。
