Research Paper Volume 13, Issue 2 pp 2436—2458

FX5 as a non-steroidal GR antagonist improved glucose homeostasis in type 2 diabetic mice via GR/HNF4α/miR-122-5p pathway

FX5 inhibited Dex or glucagon-stimulated gluconeogenesis. (A) Mouse primary hepatocytes were pretreated with Dex (10 nM) for 12 h, followed by incubation with Dex and FX5 (1, 5 and 10 μM) for 6 h in glycogenetic medium. Glucose concentration in the medium was detected to evaluate glucose output level. (B, C) Mouse primary hepatocytes were treated with Dex (10 nM) and different concentrations of FX5 (1, 5 and 10 μM) for 6 h, and mRNA levels of G6Pase and PEPCK were measured by quantitative RT-PCR analysis. (D) Glucose production assay was conducted in mouse primary hepatocytes. Cells were pretreated with glucagon (10 nM) for 12 h, and then cultured for another 6 h in glycogenetic medium with glucagon and different concentrations of FX5 (1, 5 and 10 μM). Finally, glucose level in the medium was measured. (E, F) Mouse primary hepatocytes were treated with glucagon (10 nM) and different concentrations of FX5 (1, 5 and 10 μM) for 6 h, and mRNA levels of G6Pase and PEPCK were detected by qRT-PCR analysis and normalized to GAPDH. All results were presented as mean ± S.E.M (n=3/group) (**PP

Figure 2. FX5 inhibited Dex or glucagon-stimulated gluconeogenesis. (A) Mouse primary hepatocytes were pretreated with Dex (10 nM) for 12 h, followed by incubation with Dex and FX5 (1, 5 and 10 μM) for 6 h in glycogenetic medium. Glucose concentration in the medium was detected to evaluate glucose output level. (B, C) Mouse primary hepatocytes were treated with Dex (10 nM) and different concentrations of FX5 (1, 5 and 10 μM) for 6 h, and mRNA levels of G6Pase and PEPCK were measured by quantitative RT-PCR analysis. (D) Glucose production assay was conducted in mouse primary hepatocytes. Cells were pretreated with glucagon (10 nM) for 12 h, and then cultured for another 6 h in glycogenetic medium with glucagon and different concentrations of FX5 (1, 5 and 10 μM). Finally, glucose level in the medium was measured. (E, F) Mouse primary hepatocytes were treated with glucagon (10 nM) and different concentrations of FX5 (1, 5 and 10 μM) for 6 h, and mRNA levels of G6Pase and PEPCK were detected by qRT-PCR analysis and normalized to GAPDH. All results were presented as mean ± S.E.M (n=3/group) (**P<0.01 and ***P<0.001).