Abstract
Purpose:
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.
Methods:
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.
Results:
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.
Conclusions:
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.