ALPHA-1 ANTITRYPSIN DIMINISHED ANGIOGENESIS IN HUMAN MICROVASCULAR CELLS EXPOSED TO HYPERGLYCEMIA

SALICA, JP; ORTIZ, G; GALLO, JE; CHULUYAN, E

Seattle

Congreso; ARVO Annual Meeting; 2016

ASSOCIATION FOR RESEARCH IN VISION AND OPHTHALMOLOGY

Purpose:We have previously shown that AAT diminished the activity of MMP-2 and lowered the levels of IL-6 in human microvascular endothelial cells exposed to hyperglycemia. In the present study we aim to analyze the effect of AAT on VEGF and angiogenesis.Methods:The HMEC-1 cell line were maintained in DMEM Medium supplemented with 50 and 100 mM glucose, 10% foetal bovine serum and 0.48% (w/v) HEPES, pH 7.4, in a CO2 humid incubation chamber at 37°C. The cells were exposed to 0, 1.5, and 3 mg/ml of AAT (Prolastin C®) and incubated for 3, 5 and 7 days. VEGF was measured with ELISA and angiogenesis was evaluated through the formation of capillary tubes measuring the amount of branching points.Results:VEGF increased over time for both 50mM and 100 mM glucose in control and in those treated with AAT. However, we observed that the number of branching points (BP) decreased with the use of AAT both at 5 and 7 days of culture. In particular when we used 1.5 mg/ml of AAT at 5 days the branching point diminished a third part, reaching a half part at 7 days when compared to the control condition. Furthermore, the BP diminish over time at both concentration of AAT, 1.5 and 3 mg/ml.Conclusions:AAT seems to modulate the mechanism of initial angiogenesis. Its antiangiogenic effect might be useful for the treatment of early diabetic retinopathy.