Characterization of Site-Specific Phosphorylation of NF-κ B p65 in Retinal Cells in Response to High Glucose and Cytokine Polarization

Mediators of Inflammation
Haoshen Shi, Elizabeth A Berger

Abstract

Inflammation is an important contributor to the pathogenesis of diabetic retinopathy (DR). NF-κB is a master transcriptional regulator for numerous inflammatory genes. Although NF-κB is comprised of multiple subunits, p65 has received the most attention. However, the p65 subunit can be phosphorylated at numerous sites, for which the effects of DR-related conditions are not well characterized. Since dysregulation of NF-κB has been linked to chronic inflammation, the current study examines site-specific p65 phosphorylation in retinal cells exposed to high glucose and investigates the effects of cytokine polarization. Phosphorylation of NF-κB p65 sites was examined in human primary retinal endothelial cells (HREC) and MIO-M1 Müller cells after exposure to high glucose (HG) and pro- or anti-inflammatory cytokines. Related downstream gene activation was selectively measured by real-time RT-PCR, ELISA, and/or Western blot. HG exposure resulted in differential phosphorylation of p65 subunit sites between HREC and Müller cells. Proinflammatory cytokines further increased phosphorylation of these sites and additional sites that were not altered in HG. In contrast, IL-4 exhibited a suppressive effect on the phosphorylation of p65 sites i...Continue Reading

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Citations

Nov 25, 2020·Acta Neuropathologica Communications·Haoshen ShiMaya Koronyo-Hamaoui
May 1, 2021·The Journal of Biological Chemistry·P Vineeth DanielProsenjit Mondal

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