Inducible nitric oxide synthase mediates hypoxia-induced hypoxia-inducible factor-1 alpha activation and vascular endothelial growth factor expression in oxygen-induced retinopathy.

OBJECTIVE Previous studies provided evidence that many factors contribute to retinal angiogenesis, including inducible nitric oxide synthase (iNOS), hypoxia-inducible factor-1 alpha (HIF-1 alpha) and vascular endothelial growth factor (VEGF). But the role of nitric oxide generated by iNOS in the regulation of expression of hypoxia-inducible genes in retinopathy of prematurity remains unclear. So we sought to better define the molecular basis of this iNOS-dependent regulation. METHODS In this study, using immunohistochemistry, real-time PCR and Western blotting technologies, we investigated the changes of iNOS, HIF-1 alpha, VEGF and phosphatidylinositol 3-kinase/Akt (PI3K/Akt) expressions. RESULTS Hypoxia- induced overexpression of iNOS, HIF-1 alpha, VEGF, PI3K/Akt and phosphorylated PI3K/Akt was observed in the untreated retinopathy of the prematurity group. Administration of the selective iNOS inhibitor aminoguanidine hemisulfate markedly decreased the expression of these genes. CONCLUSIONS These results indicate that iNOS mediates HIF-1 alpha activation and VEGF expression in retinal angiogenesis and that the PI3K/Akt signaling pathway may play a role in this process.