Abstract
Purpose
Müller cells have important roles in the pathogenesis of diabetic retinopathy by promoting cell proliferation and inducing the production of vascular endothelial growth factor (VEGF) under hyperglycemic conditions. The objective of this study was to determine the potential mechanism of Müller cell proliferation and VEGF production due to high-glucose conditions.
Methods
Primary cultured rat Müller cells were incubated with medium containing variable concentrations of glucose and/or embelin, a specific inhibitor of X-linked inhibitor of apoptosis protein (XIAP), for 72 h. The proliferation of Müller cells was assessed by the MTT assay. The expression and/or phosphorylation of 146 proteins were assessed using protein pathway array.
Results
High concentrations of glucose-induced Müller cell proliferation and altered expression and/or phosphorylation of 47 proteins that have been identified to have key roles in several important signaling pathways (XIAP, VEGF, HIF1α, NFκB, etc) and are involved in the regulation of cell survival, proliferation, or apoptosis. However, Müller cell alterations induced by high-glucose conditions were counteracted by the XIAP inhibitor embelin, and 26 proteins/phosphorylations (out of 47) were restored to their normal levels. Nine proteins, including NFκB p65, p-p38, tumor necrosis factor-α, urokinase-type plasminogen activator, CREB, IL-1β, HCAM, estrogen receptor-α, and p-Stat3, were involved in regulatory networks between XIAP and VEGF.
Conclusions
The current study suggests that XIAP may be a potential regulator that can mediate a series of pathological changes induced by high-glucose conditions in Müller cells. Therefore, embelin could be a potential agent for the prevention and treatment of diabetic retinopathy.
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Acknowledgements
This study was partially supported by a grant from the National Science Foundation (No. 81070736) to E Song. The authors are grateful for Drs Jianhua Liu and Boxun Xie for their kind technical support.
Author contributions
DZ, ES, and FY contributed equally to this work by conceiving the hypothesis and overall study design. YS, DZ, and FY contributed equally to the experimental design, and DW and YS contributed equally by performing the protein pathway array assay. YS, XL, and LZ contributed equally to cell preparation. LG and YS contributed equally to the protein preparation. YS, DW and FY contributed equally to statistical analysis, and YS and DW contributed equally to summarizing the final data. YS and DZ contributed equally in preparing the manuscript.
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Sun, Y., Wang, D., Ye, F. et al. Elevated cell proliferation and VEGF production by high-glucose conditions in Müller cells involve XIAP. Eye 27, 1299–1307 (2013). https://doi.org/10.1038/eye.2013.158
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DOI: https://doi.org/10.1038/eye.2013.158
