Dietary flavonoid apigenin inhibits high glucose and tumor necrosis factor α-induced adhesion molecule expression in human endothelial cells Kazuo Yamagata,

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Dietary flavonoid apigenin inhibits high glucose and tumor necrosis factor α-induced adhesion molecule expression in human endothelial cells Kazuo Yamagata, Akinori Miyashita, Hiroshi Matsufuji, Makoto Chino Journal of Nutritional Biochemistry Volume 21, Issue 2, Pages 116-124 (February 2010) DOI: 10.1016/j.jnutbio.2008.11.003 Copyright © 2010 Elsevier Inc. Terms and Conditions

Fig. 1 Effects of incubation times on HG- and TNFα-induced ICAM1, MCP1 and VCAM1 gene expression in human EC. A, ICAM 1; B, MCP 1; C, VCAM 1. The human EC line ISO-HAS was cultured for 2, 4 and 8 h with 5.5 mM glucose, HG (30 mM) and TNFα (10 ng/ml). The data are expressed as mean±S.D (n=4). Journal of Nutritional Biochemistry 2010 21, 116-124DOI: (10.1016/j.jnutbio.2008.11.003) Copyright © 2010 Elsevier Inc. Terms and Conditions

Fig. 2 Effects of flavonoid concentrations on ICAM1, MCP1 and VCAM1 gene expression in human EC. The human EC line ISO-HAS was cultured for 4 h without and with 10, 30 and 50 μM apigenin (A), caffeic acid (B), chrysin (C) and quercetin (D), and then challenged with HG (30 mM) and TNFα (10 ng/ml). The structures of apigenin, caffeic acid, chrysin and quercetin are indicated (E). The data are expressed as mean±S.D (n=4). *P<.05, significantly different from 0 μM. **P<.01, significantly different from 0 μM. Journal of Nutritional Biochemistry 2010 21, 116-124DOI: (10.1016/j.jnutbio.2008.11.003) Copyright © 2010 Elsevier Inc. Terms and Conditions

Fig. 3 Inhibitory effects of apigenin against adhesion of U937 cells by cultured human EC. The human EC line ISO-HAS was cultured for 24 h without and with 50 μM apigenin and then challenged with HG (30 mM) and TNFα (10 ng/ml). The U937 cells (1×106 cells/ml) were added and incubated with EC at 37°C for 3 h. The adhesion of U937 cells to EC was measured using quantitative monolayer adhesion assays, similar to those previously described [27]. Glucose 5.5 mM (A); glucose 30 mM (B); glucose 30 mM+TNFα 10 ng/ml (C); and glucose 30 mM+TNFα 10 ng/ml+apigenin 50 μM (D). The inhibitory effects of apigenin are indicated (fold) (E). The number of bound U937 cells per square millimeter was determined by direct microscopy. The data are expressed as mean±S.D (n=6). Journal of Nutritional Biochemistry 2010 21, 116-124DOI: (10.1016/j.jnutbio.2008.11.003) Copyright © 2010 Elsevier Inc. Terms and Conditions

Fig. 4 Effects of several flavonoids on IKKα and IKKɛ/IKKi gene expression in human EC. ISO-HAS cells were cultured for 4 h without and with 50 μM apigenin, caffeic acid, chrysin and quercetin, and then challenged with HG (30 mM) and TNFα (10 ng/ml). IKKα (A) and IKKɛ/IKKi (B). Ap, apigenin; Ca, caffeic acid; Chr, chrysin; Kae, kaempferol; Que, quercetin. The data are expressed as mean±S.D (n=4). *P<.05, significantly different from 30 mM glucose+10 ng/ml TNFα. **P<.01, significantly different from 30 mM glucose+10 ng/ml TNFα. Journal of Nutritional Biochemistry 2010 21, 116-124DOI: (10.1016/j.jnutbio.2008.11.003) Copyright © 2010 Elsevier Inc. Terms and Conditions