Muller cells treated with alpha 1 antitrypsin (AAT) in vitro showed less expression of VEGF and IL6

GALLO, JE; ORTIZ, G

Seattle

Congreso; ARVO Annual Meeting; 2016

ASSOCIATION FOR RESEARCH IN VISION AND OPHTHALMOLOGY

Muller cells treated with alpha 1 antitrypsin (AAT) in vitro showed less expression of VEGF and IL6Purpose: Diabetic retinopathy (DR) is in part consequence of a chronic hyperglycemic condition. Early stages of diabetic retinopathy are characterized by inflammatory and microvascular changes. It is known that Muller cells are responsible for secrete several molecules involved in angiogenesis and inflammation.The role of several endogenous anti-inflammatory molecules and how these interact with the endothelium in this disease is still under discussion. The aim of our work was evaluate the levels of Interleukin 6 (IL6) and vascular endothelial growth factor (VEGF) in Muller cells cultured with or without AAT.Methods: Muller cells were obtained from retinas of c57/BL6 mice. In order to characterize the isolated cells were used to perform an immunofluorescence assay with antibodies against GFAP and Vimentin. Cells were growth in DMEM Medium (Invitrogen) supplemented with 30, 50, and 100 mM glucose, 10% fetal bovine serum (FBS) 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, 3 and 4.5 mg/ml of AAT (prolastin C®) and incubated for 3 or 5 days. For metabolic control we perform MTT assay. The mRNA of il6 and tnf- alpha were measured by real time PCR and expression of VEGF and IL6 were evaluated with Enzyme Linked Immunoassay (ELISA).Results: We observed a decrease expression of VEGF in Muller cells treated with AAT (1.5, 3, 4.5mg/ml) exposed to 30 and 100mM of glucose (p 0.01) at 5 days. Besides, we observed a less expression of IL6 in supernatant of Muller cells at 100mM of glucose at 3 days at the same concentrations of AAT.Conclusions: We thought that these findings help us to a better understanding about the inflammatory process associated with stress induced for a high amount of glucose as occur in DR. Also, the use of AAT as anti-inflammatory could be a promising molecule to approach this disease.