Figure 3.

Glucose production and oxygen consumption in metformin-exposed fetal and juvenile hepatocytes. Expression of major gluconeogenic genes, PCK1 and G6PC, in fetal (n = 4) and postnatal (n = 8) Japanese macaque liver tissue (A). Glucose production was measured in the presence of substrates (lactate, pyruvate) and in response to metformin at 0-, 250-, and 1,000-μmol/L doses in fetal sheep (n = 9 hepatocyte preparations) (B), fetal rhesus macaque (n = 3) (C), and juvenile Japanese macaque (n = 4) (D) hepatocytes. G6PC gene expression was also measured in hepatocytes that were treated with 0, 250, and 1,000 μmol//L metformin for 24 h (n = 3 or 4, as shown). A mitochondrial stress test was performed, and oxygen consumption rates (OCR) were measured by Seahorse XF analysis using concentrations of 2 μmol/L oligomycin, 1 μmol/L FCCP, and 5 μmol/L rotenone with 10 μmol/L antimycin A in fetal rhesus macaque hepatocytes (E). Basal and maximal OCR and ATP production were measured in response to metformin exposure (n = 6) (F). Each experiment was performed in three to nine sets of hepatocytes isolated from different animals. For gene expression, the connected lines and dots represent the mean of treatment duplicates within a set of hepatocytes (from one animal), and results are relative to the average of the basal treatment across all sets of hepatocytes. Means ± SE are shown. *P < 0.05 compared with basal.