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.