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K. Liu, X. Xu, W. Ye, H. Liu, J. Jiang; The Role of N-Glycosylation in High Glucose-Induced Upregulation of Intercellular Adhesion Molecule-1 on Bovine Retinal Endothelia Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1358.
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© ARVO (1962-2015); The Authors (2016-present)
Chronic inflammation has been implicated the development of diabetic retinopathy. Given the definite role of intercellular adhesion molecule-1 (ICAM-1) in the inflammatory course, the effect of N-glycosylation on the upregulated expression of ICAM-1 at the surface of bovine retinal endothelial cells (BRECs) induced by high concentrations of glucose was investigated.
The gene and protein expression of ICAM-1 in primary BRECs cultured in medium containing increasing concentrations of glucose in the presence or absence of tunicamycin were studied using reverse transcription polymerase chain reaction and Western blotting analysis, and the expression level of ICAM-1 on the surface of BRECs was examined using flow cytometry. Lectin blot assay with phaseolus vulgaris leucoagglutinin (PHA-L) was performed to explore the level of N-glycans on cell total proteins or immunoprecipitated ICAM-1 with or without exposure to high glucose.
The mRNA and protein levels of ICAM-1, as well as the level of ICAM-1 at cell surface, were significantly upregulated by increasing concentrations of glucose in culture medium, with a peak at 20 mM. Consistent with this, a dramatic increase in the N-glycosylation of ICAM-1 in BRECs cultured with high concentrations of glucose was observed, which was attenuated by the treatment with tunicamycin.
High glucose-induced upregulation of ICAM-1 on the surface of BRECs was associated with increased N-glycosylation of ICAM-1. The N-glycosylation of ICAM-1 may be potential target for management of diabetic retinopathy.
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