Fig. 4: Inhibiting 11β-HSD1 activity restores impaired osteogenic activity and glucose uptake in osteoblasts with GC signaling overactivation.
From: Targeting osteoblastic 11β-HSD1 to combat high-fat diet-induced bone loss and obesity
a The mRNA expression of glucocorticoid target gene Glucocorticoid-induced leucine zipper (Gilz) in MC3T3-GFP cells and MC3T3-HSD1 cells after treating with 11β-HSD1 inhibitor (AZD8329). b, c The mRNA and protein expression of early growth response 2 (Egr2) in MC3T3-GFP cells and MC3T3-HSD1 cells after treatment of AZD8329. b The mRNA expression. c The protein expression. d The mRNA expression of Egr2 in MC3T3-GFP cells and MC3T3-HSD1 cells after treating with AZD8329 during osteogenic differentiation. e–h The osteogenic activity of MC3T3-GFP cells and MC3T3-HSD1 cells after treatment of AZD8329. e Alizarin red staining. f Bone gamma-carboxyglutamate protein (Bglap) mRNA expression. g Runt-related transcription factor 2 (Runx2) mRNA expression. h Osterix mRNA expression. i–j The glucose uptake test and mRNA expression levels of Glucose transporter type 4 (Glut4) and Phosphatidylinositol-3-kinase catalytic subunit beta (Pik3cb) in MC3T3-GFP cells and MC3T3-HSD1 cells after treatment of AZD8329. g: Glucose uptake test. h Glut4 and Pik3cb mRNA expression. Note: Data were presented as mean value ± SEM for (a, b, d–j). n = 3 biologically independent samples for osteogenic staining, western blot analysis and LC-MS/MS analysis, n = 6 biologically independent samples for RT-qPCR analysis. Statistical significance was calculated using one-way ANOVA followed by Tukey’s post-hoc test (a–c), and two-way ANOVA followed by a two-stage step-up method by Benjamini, Krieger and Yekutieli (d–j) to adjust for multiple comparisons. All tests were two-sided.
