Protective effect of diphenyleneiodonium, a NADPH oxidase inhibitor, on hyperpermeability of endothelial cells exposed to AOPP-HSA in vitro / 中国病理生理杂志

ABSTRACT

[ ABSTRACT ] AIM: To investigate the effect of advanced oxidation protein product-human serum albumin ( AOPP-HSA) at different concentrations on the permeability of human umbilical vein endothelial cell ( HUVEC) monolayer and the protective effect of NADPH oxidase inhibitor diphenyleneiodonium ( DPI ) against AOPP-HSA exposure. METHODS: Cultured HUVECs were exposed to 200 mg/L HSA (control) or AOPP-HSA (50, 100 and 200 mg/L).The permeability of the endothelial monolayer was assessed by measuring CMFDA-labeled THP-1 cells across the endothelial cells.The cultured HUVECs were treated with HSA (200 mg/L), AOPP-HSA (200 mg/L), or AOPP-HSA (200 mg/L)+DPI (100 μmol/L), and the activation of NADPH oxidase, endothelial monolayer permeability and cytoskeleton rear-rangement were evaluated.RESULTS: AOPP-HSA increased the permeability of the endothelial cell monolayer, and AOPP-HSA at 200 mg/L significantly increased the phosphorylation level of NADPH oxidase in the cells.Treatment with 100 μmol/L DPI obviously attenuated AOPP-HSA-induced NADPH oxidase activation, the increase in the permeability of the cell monolayer and the cytoskeleton rearrangement.CONCLUSION: AOPP-HSA increases the hyperpermeability of HUVEC monolayer via the phosphorylation of NADPH oxidase, and the NADPH oxidase inhibitor DPI reverses such effects.