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Gustavo Ortiz, Jorge Mancini, Juan E Gallo; Alpha-1-Antitrypsin Reduced IL-6 mRNA Levels in Human Microvascular Cells Exposed to a High Amount of Glucose. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5180.
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© ARVO (1962-2015); The Authors (2016-present)
Diabetic retinopathy is in part consequence of a chronic hyperglycemic condition. Early stages of diabetic retinopathy are characterized by inflammatory and microvascular changes. The role of several endogenous anti-inflammatory molecules and how these interact with the endothelium in this disease is still under discussion. We aimed at evaluating the effect of A1AT in human microvascular endothelial cells.
The HMEC-1 cell line were maintained in DMEM Medium (Invitrogen) supplemented with 30, 50, and 100 mM glucose, 10% foetal 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, and 3 mg/ml of AAT (Prolastin C®) and incubated for 3, 5 and 7 days. For metabolic control we perform MTT assay. The mRNA of IL-6 was measured, and zymography assay for MMP-9 and MMP-2 was performed.
We found a decrease activity of MMP-2 in cells with 30mM and 50mM of glucose treated with 1.5 and 3 mg/ml of AAT, but we did not see a correlation of MMP-9 activity and each condition. There was a relationship between mRNA of IL-6 and glucose levels. Increased glucose levels were accompanied by a rise in transcript levels of IL-6. Furthermore, IL-6 mRNA level was found lower using 3mg/ml of AAT compared with 1.5mg/ml in cells exposed to 100mM of glucose.
These findings might help to get a better understanding of the inflammatory process associated with stress induced by a high amount of glucose. In addition, the use of AAT seems to be an interesting anti-inflammatory agent to test in diabetic retinopathy.
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