Hypoxia Inducible Factor–Dependent Regulation of Angiogenesis by Nitro–Fatty Acids

OBJECTIVE—Nitro-fatty acids (NO2-FAs) are emerging as a new class of cell signaling mediators. Because NO2-FAs are found in the vascular compartment and their impact on vascularization remains unknown, we aimed to investigate the role of NO2-FAs in angiogenesis. METHODS AND RESULTS—The effects of nitrolinoleic acid and nitrooleic acid were evaluated on migration of endothelial cell (EC) in vitro, EC sprouting ex vivo, and angiogenesis in the chorioallantoic membrane assay in vivo. At 10 μmol/L, both NO2-FAs induced EC migration and the formation of sprouts and promoted angiogenesis in vivo in an NO-dependent manner. In addition, NO2-FAs increased intracellular NO concentration, upregulated protein expression of the hypoxia inducible factor-1α (HIF-1α) transcription factor by an NO-mediated mechanism, and induced expression of HIF-1α target genes, such as vascular endothelial growth factor, glucose transporter-1, and adrenomedullin. Compared with typical NO donors such as spermine-NONOate and deta-NONOate, NO2-FAs were slightly less potent inducers of EC migration and HIF-1α expression. Short hairpin RNA–mediated knockdown of HIF-1α attenuated the induction of vascular endothelial growth factor mRNA expression and EC migration stimulated by NO2-FAs. CONCLUSION—Our data disclose a novel physiological role for NO2-FAs, indicating that these compounds induce angiogenesis in an NO-dependent mechanism via activation of HIF-1α.