Effect of vasoconstrictor coupling factor 6 on gene expression profile in human vascular endothelial cells: enhanced release of asymmetric dimethylarginine
Effect of vasoconstrictor coupling factor 6 on gene expression profile in human vascular endothelial cells: enhanced release of asymmetric dimethylarginine
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DOI：
10.1097/01.hjh.0000209985.66853.1e
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发表时间：
2006-03-01
影响因子：
4.9
通讯作者：
Okumura, K
中科院分区：
文献类型：
--
作者：
Tanaka, M;Osanai, T;Okumura, K
偶联因子6（CF 6）是ATP合成酶的一种成分，抑制磷脂酶A（2）并诱导血管收缩。然而，由于花生四烯酸在血管生物学的广泛领域发挥作用，因此除了血管收缩之外，CF 6还可能发挥深远的作用。方法和结果用cDNA微阵列（n = 3）检测10（-7）mol/l的CF 6暴露24 h后，包括neuregulin-1、neuregulin-2、neuregulin-3、neuregulin-4、neuregulin-5、neuregulin-6、neuregulin-7、neuregulin-8、neuregulin-9、neuregulin-8、neure（1.84 ± 0.07倍，与对照相比，P < 0.05）和松弛素-1（1.74 ± 0.20，P < 0.05），均与充血性心力衰竭、尿激酶型纤溶酶原激活物受体有关（1.77 ± 0.24，P = 0.06）和雌激素受体P（1.74 ± 0.36，P = 0.08），两者均与血管炎症和细胞浸润有关;蛋白质精氨酸甲基转移酶（PRMT-1; 1.73 ± 0.20，P < 0.05）。在这些基因中，与不对称二甲基精氨酸（ADMA）（一氧化氮合酶（NOS）的内源性抑制剂）的合成相关的酶（PRMT-1）与降解酶二甲基精氨酸二甲氨基水解酶2（DDAH-2）一起被进一步检查。通过实时定量逆转录-聚合酶链反应测定的PRMT-1与甘油醛3-磷酸脱氢酶（GAPDH）mRNA的比率增加了9 +/-2%（n = 10，P < 0.01），而DDAH-2与GAPDH的比值降低了12 +/- 2%（n = 8，P < 0.01）。CF 6使DDAH-2蛋白和活性分别降低28 ± 5%（n = 5，P < 0.01）和19.2%（n = 6，P < 0.01）。CF 6使ADMA释放增加20 ± 8%，使NOS活性降低13 ± 1%（均为n = 8，P < 0.05）。结论CF 6通过促进ADMA的合成和抑制其降解，改变了ADMA的基因表达谱，具有致动脉粥样硬化的作用。
Background Coupling factor 6 (CF6), a component of ATP synthase, inhibits phospholipase A(2) and induces vasoconstriction. However, because arachidonic acid acts in the widespread fields of vascular biology, CF6 might exert profound effects in addition to vasoconstriction. We investigated the effect of CF6 on the gene expression profile in human umbilical vein endothelial cells.Methods and results The increased gene expression after 24-h exposure to CF6 at 10(-7) mol/l, assessed by cDNA microarray (n = 3), included neuregulin-1 (1.84 +/- 0.07 fold compared with control, P < 0.05) and relaxin-1 (1.74 +/- 0.20, P < 0.05), both relating to congestive heart failure, urokinase type plasminogen activator receptor (1.77 +/- 0.24, P = 0.06) and estrogen receptor P (1.74 +/- 0.36, P = 0.08), both relating to vascular inflammation and cell infiltration, and protein arginine methyltransferase (PRMT-1; 1.73 +/- 0.20, P < 0.05). Out of these genes, the enzyme relating to the synthesis (PRMT-1) of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase (NOS), was further examined concomitantly with the degradation enzyme, dimethylarginine dimethylaminohydrolase 2 (DDAH-2). The ratio of PRMT-1 to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA, measured by real-time quantitative reverse transcription-polymerase chain reaction, was increased by 9 +/- 2% (n = 10, P < 0.01) at 48 h after CF6 at 10(-7) mol/l, whereas the ratio of DDAH-2 to GAPDH was decreased by 12 +/- 2% (n = 8, P < 0.01). DDAH-2 protein and activity were decreased by 28 +/- 5% (n = 5, P < 0.01) and 19 2% (n = 6, P < 0.01) by CF6, respectively. ADMA release was enhanced by 20 +/- 8% and NOS activity was decreased by 13 +/- 1% (both n = 8, P < 0.05) by CF6.Conclusions CF6 changes the gene expression profile to be proatherogenic and functions as a novel stimulator for ADMA release by enhancing its synthesis and suppressing its degradation.