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. 2022 Sep 11;65:101598. doi: 10.1016/j.molmet.2022.101598

Figure 8.

Figure 8

Differential effects of TZDs and MSDC-0602K on cellular metabolism in AT-MSCs and BM-MSCs. 3T3-L1 were acutely treated with 30 μM concentration of different TZDs (rosiglitazone-ROSI, pioglitazone-PIO) and TZD analog (MSDC-0602), and MPC inhibitor (UK5099) and their effect on mitochondrial respiration was analyzed. (A-B) Changes in oxygen consumption rate (OCR) of 3T3-L1 cells in response to treatment with TZDs, TZD analog and UK5099 treatment and corresponding calculations of basal respiration, maximal respiration and spare capacity. Data are presented as mean ± SEM from at least two independent experiments with five replicates per condition; one-way ANOVA, Tukey's multiple comparison test, a: Veh vs other groups; b: PIO vs other groups, c: ROSI vs other groups. (C-D) Gene expression profile measured in (C) hBM-MSC cells (PPARG, ADIPOQ, MPC1, MPC2, GLUT4) and (D) 3T3-L1 cells (Pparγ2, Adipoq, Mpc1, Mpc2, Glut4) differentiated to AD treated with 1 μM PIO, ROSI and MSDC-0602K. Data are presented as mean ± SEM (n = 3 per condition), one-way ANOVA Tukey's multiple comparison test, a: Veh vs other groups; b: PIO vs other groups; c: ROSI vs other groups. (E-F) Gene expression profiling of enzymes involved in glutamine metabolism measured in (E) hBM-MSCs (SLC1A5, GLS, GSS) and (F) 3T3-L1 cells (Slc1a5, Gls, Gls2). Data are presented as mean of F.C. ± SEM (n = 3–5 per condition); t-test, ∗p < 0.05: PIO vs MSDC; (G) Measurement of intracellular glutamine in 3T3-L1 cells after 3 h incubation with 1 μM INS, 30 μM PIO/MSDC. Data are presented as the fold change of normalized RLU value/protein of stimulated cells over non-stimulated cells. Data are presented as mean ± SEM (n = 5 per group); ordinary one-way ANOVA, Tukey's multiple comparison test, a: INS vs other groups, b: PIO vs other groups; (H) Measurement of glucose uptake in 3T3-L1 cells after 1.5-hour incubation with 1 μM INS, 30 μM PIO/ROSI/MSDC and 2 μM UK5099. Data are presented as the fold change of normalized RLU value/protein of stimulated cells over non-stimulated cells. Data are presented as mean ± SEM (n = 5–7 per group); ordinary one-way ANOVA, Tukey's multiple comparison test, a: INS vs other groups, b: PIO vs other groups.