Figure 4.

Effects of metformin on iron overload in vitro NAFLD model

(A) Concentrations of total iron, Fe²⁺and Fe³⁺ in pellets of WRL68 cells treated with PA (0.4 mM), and FAC (0.1 mM) without or with metformin (2.5 mM) or AICAR (1.0 mM). Data are represented as mean ± SEM (n = 3 in each group). Differences among groups were assessed by one-way ANOVA analysis. ^∗∗p < 0.01, vs. control group; ^$p < 0.05, ^$$p < 0.01, vs. PA + FAC group.

(B) Concentrations of total iron, Fe²⁺ and Fe³⁺ in supernatants of WRL68 cells treated with PA (0.4 mM), and FAC (0.1 mM) without or with metformin (2.5 mM) or AICAR (1.0 mM). Data are represented as mean ± SEM (n = 3 in each group). Differences among groups were assessed by one-way ANOVA analysis. ^∗p < 0.05, ^∗∗p < 0.01, vs. control group; ^$p < 0.05, ^$$p < 0.01, vs. PA + FAC group; ^&&p < 0.01, vs. PA+FAC+Metformin group.

(C and D) Intracellular levels of TG and TC in WRL68 cells treated with FAC (0.1 mM, 0.2 mM) without or with PA (0.4 mM) in the absence or presence of metformin (2.5 mM). Data are represented as mean ± SEM (n = 3 in each group). Differences among groups were assessed by one-way ANOVA analysis. ^∗∗p < 0.01, vs. control group; ^##p < 0.01, vs. FAC 0.2 mM group; ^$$p < 0.01, vs. PA + FAC 0.1 mM group; ^&&p < 0.01, vs. PA + FAC 0.2 mM group.

(E–G) Intracellular levels of MDA, GSH and T-SOD in WRL68 cells treated with FAC (0.1 mM, 0.2 mM) without or with PA (0.4 mM) in the absence or presence of metformin (2.5 mM). Data are represented as mean ± SEM (n = 3 in each group). Differences among groups were assessed by one-way ANOVA analysis. ^∗∗p < 0.01, vs. control group; ^$$p < 0.01, vs. PA + FAC group; ^&&p < 0.01, vs. PA+FAC+Metformin group. TG, triglyceride; TC, total cholesterol; FAC, ferric ammonium citrate; MDA, malondialdehyde; GSH, glutathione; T-SOD, total superoxide dismutase.