Figure 3.

Effect of lipopolysaccharide (LPS) on iron homeostasis and renal Epo mRNA expression. C57BL/6J mice were injected intraperitoneally (i.p.) with saline (0.9% NaCl, indicated as Vehicle) or LPS (50 mg/kg). Samples were collected at 0, 1, 2, 4, 6, 12, and 24 hours (h) after treatment. (A) Quantitative real-time polymerase chain reaction (qRT-PCR) for hepcidin expression in liver. (B) Serum iron and (C) serum transferrin saturation. (D and E) qRT-PCR for ferroportin (Fpn) expression in (D) liver and (E) spleen. (F) Iron content in spleen. (G) Spleen weight normalized to the body weight of the animals. (H) qRT-PCR for lipocalin (Lcn2) in liver. (I) qRT-PCR for renal Epo mRNA expression. Data are expressed as fold change (2-DDCt) relative to housekeeping genes Gapdh or Hprt. Samples were measured in duplicates (vehicle, n=3-4; LPS, n=5-8). Data are represented as mean±standard deviation. All data were analyzed for normality with Shapiro-Wilk test and equivalence of variance using Levene’s test. The samples not in normal distribution were analyzed with non-parametric Kruskal-Wallis test (A and B, D-I). When the samples showed normal distribution, two-way ANOVA was performed with Bonferroni’s multiple comparison test in each vehicle- or LPS-treated group compared to 0 h (C). ns: not significant, *P<0.05, **P<0.01, ***P<0.001 compared to 0 h.