Glycated Human Serum Albumin Induces Interleukin 8 mRNA Expression through Reactive Oxygen Species and NADPH Oxidase-Dependent Pathway in Monocyte-Derived U937 Cells

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  • Higai Koji
    Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University
  • Sano Risa
    Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University
  • Satake Misaki
    Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University
  • Azuma Yutaro
    Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University
  • Matsumoto Kojiro
    Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University

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説明

Glycated human serum albumin (Glc-HSA) has previously been reported (Higai K., et al., 2006) to induce E-selectin expression on human umbilical vein endothelial cells through activation of NADPH oxidase; however, Glc-HSA signaling in monocytes remains obscure. To clarify the influence on human monocyte-derived U937 cells, U937 cells were stimulated with Glc-HSA and glycoaldehyde-dimer-modified HSA (GA-HSA) for 2 h in the absence and presence of the protein kinase C (PKC) inhibitor calphostin and the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC) and NADPH oxidase inhibitor apocynin; interleukin-8 (IL-8) mRNA expression was determined by RT-PCR. As a result, IL-8 mRNA expression in U-937 cells was time- and dose-dependently enhanced by stimulation with Glc-HSA and GA-HSA. Furthermore, promoter activity of the IL-8 reporter gene was enhanced approximately 2-fold by stimulation with Glc-HSA and GA-HSA. Nuclear factor κB (NFκB) and activator protein-1 (AP-1) reporter genes were also enhanced although CCAAT/enhancer binding protein (C/EBP) was not affected. IL-8 mRNA expression was suppressed by NAC and apocynin but not calphostin in cells stimulated with Glc-HSA; however, its expression in cells stimulated with GA-HSA was suppressed by calphostin but not NAC. These results indicated that IL-8 mRNA expression was upregulated by NFκB and AP-1 in U937 cells stimulated with Glc-HSA and GA-HSA, but the signaling pathways were different.

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