Fang Yuan1, Shang-xi Liu, Jian-wei Tian. 1. Department of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Abstract
OBJECTIVE: To investigate the effect of advanced oxidation protein products (AOPP) on production of reactive oxygen species (ROS) in endothelial cells. METHODS: Human umbilical vein endothelial cell line ECV304 were stimulated with AOPP-modified bovine serum albumin (AOPP-BSA) prepared by oxidation of BSA with HOCl in vitro. ROS production in the cells was evaluated by kinetic measurement of dichlorofluoroscein (DCF) fluorescence produced by oxidation of an oxidant-sensitive dye 2,7-dichlorefluorescin (DCFH) using VICTOR 1420 multilabel counter. RESULTS: AOPP-BSA could induce ROS production in ECV304 cells time- and dose-dependently, and the quantity of ROS increased with the oxidation degree of BSA. Pretreatment of cells with a small-molecular-mass glutathione peroxidase mimic ebselen or NADPH oxidase inhibitor apocynin inhibited ROS production by 65% and 29% respectively. CONCLUSION: AOPP-BSA induces ROS production in endothelial cells, partially mediated by NADPH oxidase activation.
OBJECTIVE: To investigate the effect of advanced oxidation protein products (AOPP) on production of reactive oxygen species (ROS) in endothelial cells. METHODS:Human umbilical vein endothelial cell line ECV304 were stimulated with AOPP-modified bovine serum albumin (AOPP-BSA) prepared by oxidation of BSA with HOCl in vitro. ROS production in the cells was evaluated by kinetic measurement of dichlorofluoroscein (DCF) fluorescence produced by oxidation of an oxidant-sensitive dye 2,7-dichlorefluorescin (DCFH) using VICTOR 1420 multilabel counter. RESULTS: AOPP-BSA could induce ROS production in ECV304 cells time- and dose-dependently, and the quantity of ROS increased with the oxidation degree of BSA. Pretreatment of cells with a small-molecular-mass glutathione peroxidase mimic ebselen or NADPH oxidase inhibitor apocynin inhibited ROS production by 65% and 29% respectively. CONCLUSION: AOPP-BSA induces ROS production in endothelial cells, partially mediated by NADPH oxidase activation.