Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells
Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells
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DOI：
10.1161/01.atv.0000219615.88323.b4
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发表时间：
2006-07-01
影响因子：
8.7
通讯作者：
Hayashi, Koichi
中科院分区：
文献类型：
--
作者：
Hasegawa, Kazuhiro;Wakino, Shu;Hayashi, Koichi
目的-二甲基精氨酸二甲基氨基水解酶（DDAH）是内源性NO合成酶抑制剂不对称二甲基精氨酸的降解酶。DDAH诱导的血管内皮生长因子（VEGF）的表达的分子机制examined.Methods和结果-虽然DDAH，DDAH 1，或DDAH 2的2种亚型的表达载体的转染增加DDAH在牛主动脉内皮细胞和人脐静脉内皮细胞的活性，表达和分泌的VEGF增加，只有在DDAH 2转染细胞。敲除DDAH 2基因减少VEGF的产生，DDAH 2过表达增强内皮细胞的增殖和迁移。VEGF启动子活性增加DDAH 2转染，这是不封闭的一氧化氮合酶（NOS）抑制剂，但需要的Sp1位点。DDAH 2过表达增加内皮细胞中与Sp1寡核苷酸结合的核蛋白水平。Sp1小干扰RNA阻断DDAH 2诱导的VEGF上调。DDAH 2转染以蛋白激酶A（PKA）依赖的方式增加Sp1的核和苏氨酸磷酸化水平。结论DDAH 2通过Sp1依赖性和NO/NOS系统非依赖性的启动子激活上调VEGF的表达。DDAH 2增加Sp1 DNA结合活性是PKA依赖性的。这些机制可能为VEGF相关的血管病变如动脉粥样硬化提供一种新的治疗策略。
Objectives - Dimethylarginie dimethylaminohydrolase (DDAH) is a degrading enzyme for asymmetrical dimethylarginine, an endogenous NO synthase inhibitor. The molecular mechanism for DDAH-induced vascular endothelial growth factor (VEGF) expression was examined.Methods and Results - Although the transfection of expression vectors for 2 isoforms of DDAH, DDAH1, or DDAH2 increased DDAH activity in bovine aortic endothelial cells and human umbilical vein endothelial cells, expression and secretion of VEGF were increased only in DDAH2-transfected cells. Knocking down the DDAH2 gene reduced VEGF production, and DDAH2 overexpression enhanced both proliferation and migration of endothelial cells. The VEGF promoter activity was increased by DDAH2 transfection, which was not blocked by an NO synthase (NOS) inhibitor but required the Sp1 sites. DDAH2 overexpression increased nuclear protein levels bound to Sp1 oligonucleotides in endothelial cells. Sp1 small interfering RNA blocked DDAH2-induced upregulation of VEGF. DDAH2 transfection increased nuclear and threonine-phosphorylation levels of Sp1 in a protein kinase A (PKA)-dependent manner. Protein - protein interaction between DDAH2 and PKA was enhanced in DDAH2-transfected cells.Conclusions - DDAH2 upregulated the expression of VEGF through Sp1-dependent and NO/NOS system-independent promoter activation. DDAH2-increased Sp1 DNA binding activity was PKA dependent. These mechanisms may provide a novel therapeutic strategy for VEGF-related vasculopathies such as atherosclerosis.