β-Glucan attenuates inflammatory responses in oxidized LDL-induced THP-1 cells via the p38 MAPK pathway  S. Wang, H. Zhou, T. Feng, R. Wu, X. Sun, N. Guan, L. Qu, Z. Gao, J. Yan, N. Xu, J. Zhao, C. Qi  Nutrition, Metabolism and Cardiovascular Diseases  Volume 24, Issue 3, Pages 248-255 (March 2014) DOI: 10.1016/j.numecd.2013.09.019 Copyright © 2013 Elsevier B.V. Terms and Conditions

Figure 1 Surface marker expression in oxLDL-induced PMA-differentiated THP-1 cells. THP-1 cells were cultured with PMA for 48 h. Cells were washed and cultured with RPMI 1640 containing 1% FBS and stimulated with oxLDL for another 48 h with or without WGP pre-treatment. Surface marker expression was assessed by flow cytometry. One representative histogram from 3 independent experiments with similar results is shown. The asterisk (*) indicates p < 0.05. Nutrition, Metabolism and Cardiovascular Diseases 2014 24, 248-255DOI: (10.1016/j.numecd.2013.09.019) Copyright © 2013 Elsevier B.V. Terms and Conditions

Figure 2 Cytokine secretion in oxLDL-induced PMA-differentiated THP-1 cells. THP-1 cells were cultured with PMA for 48 h. Cells were washed and cultured with RPMI 1640 containing 1% FBS and stimulated with oxLDL for another 48 h with or without WGP pre-treatment. Supernatants from cell culture medium were collected and assayed. The asterisk (*) indicates p < 0.05. Nutrition, Metabolism and Cardiovascular Diseases 2014 24, 248-255DOI: (10.1016/j.numecd.2013.09.019) Copyright © 2013 Elsevier B.V. Terms and Conditions

Figure 3 The involvement of p38 MAPK and ERK1/2 pathways in oxLDL-induced PMA-differentiated THP-1 cells. A). The oxLDL-induced phosphorylation of MAPK members. PMA-differentiated THP-1 cells were incubated with oxLDL for different time intervals. Phosphorylation of signal proteins was determined by Western blotting of whole cell lysates. β-actin was used as a loading control. B). β-glucan suppressed the oxLDL-induced phosphorylation of p38 MAPK. PMA-differentiated THP-1 cells were pre-treated with 100 μg/ml WGP for 2 h before oxLDL was added. p38 MAPK phosphorylation was determined by Western blot. β-actin was used as a loading control. These are representative of experiments conducted at least in triplicate. The gray values were evaluated by ImageJ software. The phosphorylation proteins gray scale data have been normalized to total proteins gray values, respectively. S: SB203580; U: U0126. *p < 0.05. Nutrition, Metabolism and Cardiovascular Diseases 2014 24, 248-255DOI: (10.1016/j.numecd.2013.09.019) Copyright © 2013 Elsevier B.V. Terms and Conditions

Figure 4 β-glucan inhibited cytokine production in oxLDL-induced circulating monocytes. Circulating monocytes from health donors were cultured with PMA for 48 h. Cells were washed, cultured with RPMI 1640 containing 1% FBS, and stimulated with oxLDL for another 48 h with or without WGP pretreatment. RNA from oxLDL-induced monocytes was extracted, and qRT-PCR was performed for the indicated cytokines. *p < 0.05. Nutrition, Metabolism and Cardiovascular Diseases 2014 24, 248-255DOI: (10.1016/j.numecd.2013.09.019) Copyright © 2013 Elsevier B.V. Terms and Conditions

Figure 5 β-glucan inhibited inflammatory cytokine production in human atherosclerotic plaque cells. A). Single cell suspensions prepared from atherosclerotic plaques were cultured with WGP for 3 days. On day 3, the supernatants were collected for ELISA. B). Single cell suspensions prepared from atherosclerotic plaques were cultured with WGP for 3 days. After 3 days, cellular RNA was extracted, and qRT-PCR was performed for the indicated cytokines and NOS2A. The data from WGP group were normalized to the data from PBS group, respectively. *p < 0.05. Nutrition, Metabolism and Cardiovascular Diseases 2014 24, 248-255DOI: (10.1016/j.numecd.2013.09.019) Copyright © 2013 Elsevier B.V. Terms and Conditions