Research Article

Acidic Polysaccharide from Angelica sinensis Reverses Anemia of Chronic Disease Involving the Suppression of Inflammatory Hepcidin and NF-κB Activation

Figure 5

ASP inhibits IL-6-induced hepcidin expression and STAT3 phosphorylation as well as translocation in HepG2 cells. The cells were grown for 16 hours in the presence or absence of ASP, then added IL-6 (50 ng/mL) and grown for another 8 hours. (a) Representative immunofluorescence images illustrated that ASP successfully suppressed p-STAT3 nuclear translocation induced by IL-6.The hepcidin mRNA levels were quantified by qRT-PCR. (b) The STAT3 phosphorylation in response to IL-6, SMAD4, and hepcidin protein expressions was analyzed by Western blotting. One representative blot of 3 independent experiments is shown; β-actin served as the loading control. (c) HepG2 cells were pretreated with or without ASP (200 μg/mL) for 16 hours and then stimulated with 50 ng/mL IL-6 in the presence or absence of AZD1480 (5 μM) for another 8 hours. p-JAK2 expression was analyzed by Western blotting. (d) The hepcidin mRNA levels were quantified by qRT-PCR. Values shown were the means of fold changes in Hamp gene expression relative to β-actin (2−ΔΔCt) ± SEM, . # and ## compared with the control group, and compared with the IL-6 group. Micrographs (magnification, 400x) are representative of one of three similar experiments and taken with Nikon E400.
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