Abstract
Abstract: :
Purpose:The kinetics of HIF1 in activating vascular endothelial growth factor (VEGF) expression was analysed in distinct cell lines. Methods:RF/6A rhesus monkey choroid–retinal and human umbilical vein cord (HUVEC) endothelial cells were incubated under hypoxic conditions for 1, 2, 4, and 8 hours and treated with the phosphoinositol–3 (PI–3) kinase inhibitor LY294002 (Calbiochem) or vehicle. Cells and culture media were harvested after each hypoxic timepoint. Nuclear extracts were used in 1) electrophoretic mobility shift assays (EMSA) with the hypoxia–inducible–factor–1 (HIF1) elements in the vascular endothelial growth factor (VEGF) and promoter and 2) Western Blots for HIF1 detection. VEGF release was determined by enzyme–linked immunoabsorbent assay (ELISA). Results: Conclusions:
Keywords: retinal neovascularization • hypoxia • choroid: neovascularization