CONICET | Buscador de Institutos y Recursos Humanos

Diabetic retinopathy is a leading cause of blindness in working-age population and is regarded as a microvascular complication due to the breakdown of the endothelial barrier. The retinal pigment epithelium (RPE), which is the major component of the outer blood-retinal barrier is also the key to maintain the integrity of retinal tissues. In diabetic retinopathy, cell-cell junction complexes between RPE cells are to disassemble and the leakage can be found through paracellular space. Dissociation of cultured RPE cells leads to dedifferentiation of the cells into fibroblast-like cells through epithelial to mesenchymal transition (EMT). To maintain the integrity of epithelial cells adherens junctions are critical. NFkB, TG2, mTOR, Wnt canonical pathway mediators like GSK3b and b-Catenin are proteins involved in many biological processes, such as inflammation, angiogenesis, cell adhesion, migration, survival and the EMT. HIF-1a, E-Cadherin and N-Cadherin are regulated with this pathways, where HIF-1a is related with angiogenesis and E-Cadherin and N-Cadherin are related with cellular adhesion. In this work, we explore the expression of NFkB, TGM2, mTOR, GSK3b, b-Catenin, HIF1a, N-Cadherin and E-cadherin on ARPE-19 cells exposed to high glycemia and A1AT treatment. ARPE-19 cells were incubated 16h with DMEM 5,5mM glucose (Control), DMEM 5,5mM glucose + 4.5mg/ml A1AT (Control + A1AT), DMEM 30mM glucose (Diabetic), DMEM 30mM glucose + 4.5mg/ml A1AT (Diabetic + A1AT). Cells were harvested for Western blot, for RT-qPCR or fixed for immunohistochemistry. NFkB, TGM2, AKT, pAKT, mTOR, b-Catenin, N-Cadherin and HIF1a were diminished with A1AT treatment, while, GSK3b and E-Cadherin were increased.Outcomes support the hypothesis that A1AT decrease integrin mediated signaling avoiding EMT. These results help to understand mechanisms involved in inflammatory processes in diabetic retinopathy and make A1AT as a suitable molecule to diabetic retinopathy treatment.