Nitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells
Nitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells
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DOI：
10.1161/01.atv.20.3.659
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发表时间：
2000-03-01
影响因子：
8.7
通讯作者：
Cooke, JP
中科院分区：
文献类型：
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作者：
Dulak, J;Józkowicz, A;Cooke, JP
已知血管内皮生长因子（VEGF）诱导内皮细胞释放一氧化氮（NO）。然而，NO对VEGF合成的影响尚不清楚。因此，内源性和外源性NO对大鼠血管平滑肌细胞（VSMCs）合成VEGF的影响进行了研究。使用两种体外模型：（1）通过用白细胞介素（IL）-1 β（10 ng/mL）处理刺激VSMC产生NO和（2）用含有内皮组成型NO合酶（ecNOS）cDNA的pEGNOS质粒脂质转染的VSMC。NOS抑制剂L-硝基精氨酸甲酯（L-NAME，2 ~ 5 mmol/L）和GTP环化水解酶Ⅰ抑制剂DAHP（2.5 ~ 5 mmol/L）可抑制NO的合成。用L-NAME或DAMP处理的部分细胞分别补充L-精氨酸（10 mmol/L）或四氢生物蝶呤（BH（4）; 100 μ mol/L）。此外，我们研究了硝普钠（SNP; 10和100 μ mol/L）和化学相关化合物，亚铁氰化钾和铁氰化钾，对VEGF生成的影响。IL-1 β诱导iNOS表达和NO生成，并显著上调VEGF mRNA表达和蛋白质合成。L-NAME和DAHP完全抑制NO的产生，并使IL-1 β上调的VEGF合成减少30%至40%。补充L-精氨酸或BH（4）增加了L-NAME或DAMP处理的细胞的NO生成，并且加入BH（4）增加了VEGF的合成。与对照质粒转染的细胞相比，转染pEGNOS后产生NO的细胞释放的VEGF量显著增加。通过L-NAME抑制NO的产生减少VEGF的合成。与内源性NO的作用相反，我们观察到在高浓度（10或100 μ mol/L）SNP存在下VEGF合成的抑制。化学相关的铁氰化物和亚铁氰化物的化合物，这表明硝普钠的抑制作用可能是由一个NO-独立的机制介导的模仿。结果表明，内源性NO促进VEGF的合成。内源性NO和VEGF之间的积极相互作用可能对球囊血管成形术后的内皮再生和血管生成有影响。
Vascular endothelial growth factor (VEGF) is known to induce the release of nitric oxide (NO) from endothelial cells. However, the effect of NO on VEGF synthesis is not clear. Accordingly, the effect of endogenous and exogenous NO on VEGF synthesis by rat vascular smooth muscle cells (VSMCs) was investigated. Two in vitro models were used: (1) VSMCs stimulated to produce NO by treatment with interleukin (IL)-1 beta (10 ng/mL) and (2) VSMCs lipotransfected with pKecNOS plasmid, containing the endothelial constitutive NO synthase (ecNOS) cDNA. The synthesis of NO was inhibited by N(omega)-nitro-L-arginine methyl ester (L-NAME, 2 to 5 mmol/L) or diaminohydroxypyrimidine (DAHP, 2.5 to 5 mmol/L), inhibitors of NOS and GTP cyclohydrolase I, respectively. Some cells treated with L-NAME or DAMP were supplemented with L-arginine (10 mmol/L) or tetrahydrobiopterin (BH(4); 100 mu mol/L), respectively. In addition, we studied the effect of sodium nitroprusside (SNP; 10 and 100 mu mol/L) and chemically related compounds, potassium ferrocyanide and ferricyanide, on VEGF generation. IL-1 beta induced iNOS expression and NO generation and significantly upregulated VEGF mRNA expression and protein synthesis. L-NAME and DAHP totally inhibited NO generation and decreased the IL-1 beta-upregulated VEGF synthesis by 30% to 40%. Supplementation with L-arginine or BH(4) increased NO generation by L-NAME- or DAMP-treated cells, and VEGF synthesis was augmented by addition of BH(4). The cells generating NO after pKecNOS transfection released significantly higher amounts of VEGF than cells transfected with control plasmids. Inhibition of NO generation by L-NAME decreased VEGF synthesis. In contrast to the effect of endogenous NO, we observed the inhibition of VEGF synthesis in the presence of high (10 or 100 mu mol/L) concentrations of SNP. This effect was mimicked by chemically related ferricyanide and ferrocyanide compounds, suggesting that the inhibitory effect of sodium nitroprusside may be mediated by an NO-independent mechanism. The results indicate that endogenous NO enhances VEGF synthesis. The positive interaction between endogenous NO and VEGF may have implications for endothelial regeneration after balloon angioplasty and for angiogenesis.