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K-S Kim, H-D Lee, S-I Suh; The Effects of High Glucose on Expression of Vascular Endothelial Growth Factor and Hepatocyte Growth Factor in Cultured Retinal Pigment Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4577.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:To evaluate the protein expression of vascular endothelial growth factor(VEGF) and hepatocyte growth factor(HGF) in response to high glucose concentration in human retinal pigment epithelial(RPE) cells. Methods:After the confluent adult human RPE cells(ARPE-19) were cultured in RPMI 1640 media containing three different concentration of glucose (5.5, 11 and 22 mM/L) for 3, 7 and 14 days, western blot analyses were performed for their VEGF and HGF expression. Cell pellets were lysed in lysis buffer and the cell lysates were centrifused to collect the supernatant fractions for quantifying the protein concentration. The proteins were subjected to sodium dodesyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. After the blots were incubated with primary and secondary antibody to VEGF or HGF consecutively, enhanced chemiluminescence was carried out to detect them. Results:: Incubation with different concentration of glucose increased the protein expression of VEGF and HGF in time-and concentration-dependent manner in RPE . In particular, at 14 days, the level of VEGF expression was higher in 22 mM/L glucose group than in other two groups (5.5 mM/L or 11 mM/L ). At 3 days, the level of HGF expression was higher in 22 mM/L glucose group than in other groups. Conclusion:RPE cells may participate in angiogenesis in progression of diabetic retinopathy through the production of VEGF and HGF.
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