Abstract
Abstract: :
Purpose:Diabetic patients are at increased risk for developing corneal disorders including recurrent epithelial erosion and delayed reepithelialization. The purpose of this work was to investigate the effect of hyperglycemia on cell proliferation and migration in RCEC induced by epidermal growth factor (EGF). We also examined whether the expression of PLCγ1 is affected under hyperglycemic conditions. Methods:Serum–starved, primary RCEC (∼50% confluent) were cultured with or without EGF and different concentrations of glucose. At prescribed times, the cultures were terminated, and the cells trypsinized and counted. The cell migration assay was performed using trans–well chambers. To determine changes in PLCγ1, suitable aliquots from RCEC cultured under hyperglycemic conditions were subjected to Western immunoblotting using antibody specific for PLCγ1. Results: Addition of EGF (10 ng/ml) to RCEC, cultured in a medium containing 17.5 mM glucose, caused a time–dependent increase in cell migration and proliferation. Increasing glucose concentration to 25 mM further increased cell migration and proliferation both in the EGF–treated and untreated RCEC. However, when glucose concentration was increased to 38 mM, both cell migration and proliferation were significantly decreased in RCEC. Western immunoblotting analysis revealed a small but significant decrease in PLCγ1 in RCEC cultured in 38 mM glucose. Conclusions: The data demonstrate that elevated glucose (>25 mM) adversely affects cell migration and proliferation in RCEC. Further, that these effects of hyperglycemia might be due to decreased expression of PLCγ1 in RCEC.
Keywords: cornea: epithelium • diabetes • wound healing