Nitric oxide stimulates vascular endothelial growth factor production in cardiomyocytes involved in angiogenesis
Nitric oxide stimulates vascular endothelial growth factor production in cardiomyocytes involved in angiogenesis
复制标题
DOI：
10.2170/physiolsci.rp002305
复制
发表时间：
2006-02-01
影响因子：
2.3
通讯作者：
Sato, Takayuki
中科院分区：
文献类型：
--
作者：
Kuwabara, Masanori;Kakinuma, Yoshihiko;Sato, Takayuki
背景：缺氧诱导因子(HIF)-1 α调节一系列基因的转录，包括血管内皮生长因子（VEGF），一个负责血管生成的主要基因。最近的一些研究表明，通过一氧化氮（NO）的非缺氧途径参与了HIF-1 α的激活。然而，没有直接证据表明心肌细胞通过心脏HIF-1 α的非缺氧诱导途径释放血管生成因子。因此，我们利用原代培养的大鼠心肌细胞（PRCMs），评估了一氧化氮供体s -亚硝基- n -乙酰青霉胺（SNAP）在常氧条件下通过HIF-1 α诱导VEGF的作用。方法与结果：乙酰胆碱（ACh）或SNAP处理的prcm均能产生大量NO。在正常缺氧状态下，SNAP激活了对prcm中HIF-1 α蛋白表达的诱导。磷脂酰肌醇3-激酶（PI3K）依赖性Akt磷酸化由SNAP诱导，并被PI3K抑制剂wortmannin和NO合酶抑制剂n - g -硝基- l -精氨酸甲酯（L-NAME）完全阻断。SNAP治疗也增加了prcm中VEGF蛋白的表达。此外，从snap处理的心肌细胞中提取的条件培养基磷酸化了人脐静脉内皮细胞的VEGF 2型受体（Flk-1）（与对照组相比增加了四倍，p < 0.001, n = 5）并加速了血管生成。结论：我们的研究结果表明，心肌细胞通过NO激活的非缺氧HIF-1 α诱导途径产生VEGF，导致血管生成。
Background: Hypoxia-inducible factor (HIF)-1 alpha regulates the transcription of lines of genes, including vascular endothelial growth factor (VEGF), a major gene responsible for angiogenesis. Several recent studies have demonstrated that a nonhypoxic pathway via nitric oxide (NO) is involved in the activation of HIF-1 alpha. However, there is no direct evidence demonstrating the release of angiogenic factors by cardiomyocytes through the nonhypoxic induction pathway of HIF-1 alpha in the heart. Therefore we assessed the effects of an NO donor, S-Nitroso-N-acetylpenicillamine (SNAP) on the induction of VEGF via HIF-1 alpha under normoxia, using primary cultured rat cardiomyocytes (PRCMs). Methods and Results: PRCMs treated with acetylcholine (ACh) or SNAP exhibited a significant production of NO. SNAP activated the induction of HIF-1 alpha protein expression in PRCMs during normoxia. Phosphatidlylinositol 3-kinase (PI3K)-dependent Akt phosphorylation was induced by SNAP and was completely blocked by wortmannin, a PI3K inhibitor, and N-G-nitro-L-arginine methyl ester (L-NAME), a NO synthase inhibitor. The SNAP treatment also increased VEGF protein expression in PRCMs. Furthermore, conditioned medium derived from SNAP-treated cardiomyocytes phosphorylated the VEGF type-2 receptor (Flk-1) of human umbilical vein endothelial cells (a fourfold increase compared to the control group, p < 0.001, n = 5) and accelerated angiogenesis. Conclusion: Our results suggest that cardiomyocytes produce VEGF through a nonhypoxic HIF-1 alpha induction pathway activated by NO, resulting in angiogenesis.