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. 2017 Mar 24;175(8):1217–1229. doi: 10.1111/bph.13753

Figure 5.

Figure 5

Metformin inhibits AngII‐induced HNF4α expression, TGFβ1 expression and cardiac fibrosis in an AMPK‐dependent manner. (A–B) Western blot analysis of HNF4α expression in CFs. (A) CFs were pretreated with Compound C (1 μM) for 0.5 h and then treated with metformin (1 mM) for 2 h. Afterwards, AngII (1 μM) was added for 24 h before harvesting. n = 5. Met: metformin; Cmpd C: Compound C. (B) CFs were pretreated with AICAR (0.1 mM) for 1 h and then treated with AngII (1 μM) for 24 h before harvesting, n = 6. (C) Western blot analysis of HNF4α expression in AMPKα2−/− mice treated with AngII and/or metformin. (D) Upper panel: quantitative real‐time PCR analysis of TGFβ1 mRNA expression. Lower panel: the TGFβ1 protein level was determined via elisa. (E) Upper panel: representative micrographs of Sirius red‐stained heart sections; the red areas represent collagen. Bars = 500 μm. Lower panel: the fibrotic area was quantified as the percentage of the total cardiac area. (F) Collagen I (upper panel) and collagen III (lower panel) mRNA expression was measured via real‐time PCR analysis. Bars represent means ± SEM of eight mice per group. *P < 0.05, NS = not significant. One‐way (A) or two‐way ANOVA with the Bonferroni post hoc test was used (B and D). Welch's ANOVA with post hoc Games–Howell test was used in (C, E and F).