June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Alpha-1-Antitrypsin Reduced IL-6 mRNA Levels in Human Microvascular Cells Exposed to a High Amount of Glucose
Author Affiliations & Notes
  • Gustavo Ortiz
    Ciencias Biomedicas, Universidad Austral, Pilar, Argentina
  • Jorge Mancini
    Ciencias Biomedicas, Universidad Austral, Pilar, Argentina
  • Juan E Gallo
    Ciencias Biomedicas, Universidad Austral, Pilar, Argentina
  • Footnotes
    Commercial Relationships Gustavo Ortiz, None; Jorge Mancini, None; Juan Gallo, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 5180. doi:
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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)

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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.

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