Nitric oxide inhibits matrix metalloproteinase-2 expression via the induction of activating transcription factor 3 in endothelial cells
Nitric oxide inhibits matrix metalloproteinase-2 expression via the induction of activating transcription factor 3 in endothelial cells
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DOI：
10.1124/mol.65.5.1130
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发表时间：
2004-05-01
影响因子：
3.6
通讯作者：
Wang, DL
中科院分区：
文献类型：
--
作者：
Chen, HH;Wang, DL
一氧化氮(NO)可抑制多种基质金属蛋白酶(MMPs)参与的细胞迁移。这种抑制的潜在分子机制仍然难以捉摸。内皮细胞(ECs)结构性地产生基质金属蛋白酶-2。观察NO对基质金属蛋白酶-2表达的影响。NO对内皮细胞基质金属蛋白酶-2基因表达的抑制作用呈剂量依赖性。携带内皮型一氧化氮合酶(Ad-eNOS)的腺病毒感染内皮细胞后，基质金属蛋白酶-2的表达减少。NO对基质金属蛋白酶-2表达的抑制是一种转录事件，因为一氧化氮降低了基质金属蛋白酶-2的启动子活性。酶谱分析显示，NO处理内皮细胞可抑制基质金属蛋白酶-2的分泌。对MMP2启动子(1716个碱基对)的功能分析表明，P53结合位点(-1659~-1629)是影响MMP2启动子活性的关键。激活转录因子3(ATF3)被认为是P53的转录抑制因子。NO处理的内皮细胞可诱导ATF3的表达。AdeNOS感染的内皮细胞ATF3水平持续升高。此外，内皮细胞过表达ATF3，或当ATF3激动剂(MG-132；苯甲氧基-L-亮氨基-L-亮氨基-L-亮氨酸)处理时，抑制基质金属蛋白酶-2启动子的活性。由于NO对基质金属蛋白酶-2的抑制作用，一氧化氮处理的内皮细胞抑制内皮细胞的迁移，这一现象类似于基质金属蛋白酶-2抗体或MG-132处理的内皮细胞。这些结果表明，NO抑制内皮细胞迁移至少部分是通过上调ATF3来减少其MMP2的表达。这项研究提供了一个分子基础，支持一氧化氮在内皮细胞迁移中起负调控作用的观点。
Nitric oxide ( NO) has been shown to inhibit migration of cells in which various matrix metalloproteinases ( MMPs) are involved. The underlying molecular mechanisms of this inhibition remain elusive. Endothelial cells (ECs) constitutively produce MMP-2. The effect of NO on MMP-2 expression was examined. A dose-dependent inhibition of MMP-2 mRNA level was demonstrated in ECs treated with NO. ECs infected with adenovirus carrying endothelial NO synthase ( Ad-eNOS) reduced MMP-2 expression. The inhibitory effect of NO on MMP-2 expression was a transcriptional event because NO reduced MMP-2 promoter activity. NO treatment of ECs consequently suppressed MMP-2 secretion revealed by zymographic assay. Functional analysis of MMP-2 promoter ( 1716 base pairs) indicated that the p53-binding site ( - 1659 to - 1629) was crucial for MMP-2 promoter activity. Activating transcription factor 3 (ATF3) has been reported to act as a transcriptional repressor for p53. ECs treated with NO induced ATF3 expression. Consistently, AdeNOS-infected ECs showed an increase of ATF3 level. Moreover, ECs either over-expressed ATF3 or, when treated with an ATF3 activator (MG-132; carbobenzoxy-L-leucyl-L-leucyl-L-leucinal), resulted in a repression of MMP-2 promoter activity. Because of MMP-2 suppression by NO, ECs treated with NO inhibited endothelial migration, a phenomenon similar to that of ECs treated with MMP-2 antibody or MG-132. These results indicate that NO-attenuating endothelial migration is mediated at least in part by its reduction of MMP-2 expression via the up-regulation of ATF3. This study provides a molecular basis that supports the notion that NO acts as a negative regulator in endothelial migration.