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Long Shi, xy wu; Effect of Advanced Glycation End Products on the generation of ROS in Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1374.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effect of Advanced Glycation End Products (AGE) on the generation of reactive oxygen species (ROS) in corneal epithelial cells and the underlying mechanisms in vitro.
Telomerase-immortalized human corneal epithelial cells (THCEs) were treated with AGE-modified bovine serum albumin (BSA) for various times. The production of ROS was measured with 2', 7'- dichlorofluorescein diacetate (DCFH-DA) dye assay and imaged on laser scanning confocal fluorescence microscope. NADPH oxidase activity was measured by luminescence assay. The protein levels of p22phox and Nox4 were determined by Western blot.
AGE-BSA significantly increased the generation of intracellular ROS in THCEs cells. However, the generation of intracellular ROS was completely inhibited by antioxidant N-acetylcysteine (NAC), anti-receptor of AGEs (RAGE) antibodies, or the inhibitor of NADPH oxidase. Moreover, AGE-BSAincreased NADPH oxidase activity and protein expression of NADPH oxidase subunits, p22phox and Nox4, but anti-RAGE antibodies eliminated these effects.
In conclusion, our results demonstrated that AGE-BSA increased intracellular ROS generation through NADPH oxidase activation, which may account for the delayed corneal epithelial wound healing. These results may provide better insights for understanding the mechanism of delayed healing of corneal epithelial wounds in diabetes.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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