Fig. 2.

Hyperglycemia induced 15-LOX protein expression as well as PLA2 activity. A: HRECs were treated with HG, D-glucose (30 mM), or normo-osmotic control for 5 days. Western blot was performed as described in the Research Design and Methods using 15-LOX, 5-LOX, and actin antibodies followed by densitometric analysis. Ratio of the band intensity of 15-LOX or 5-LOX relative to the actin was reported as fold increase in relation to normo-osmotic control (L-glucose), which was arbitrarily set at 1.0. B: HRECs were treated with 15-HETE or vehicle and the change in the resistance was monitored as described in Research Design and Methods using ECIS. Normalized TER for 15-HETE treatment was compared with vehicle-treated endothelial monolayer. C: Treatment of HRECs with D-glucose (30 mM) for 5 days stimulated the activity of PLA2 compared with normo-osmotic control. Data shown are the mean ± SD of three independent experiments. D: In vivo detection of phosphorylated PLA2 (pPLA2) (active form) in serial sections from human diabetic or normal retinas around blood vessel regions (yellow arrowhead), perivascular in the area of glial cells, detected by its marker, glial fibrillary acidic protein (GFAP) (white arrows), as well as in the outer segment of the photoreceptors (yellow arrow).