Fig. 6: Hyperglycemia mediates KC ROS production by inactivating AMPK and activating PI3K/AKT signaling pathway.

Mice were subjected to STZ pretreatment and APAP administration as described in “Materials and methods” section. KCs were isolated from different experimental groups. a p-AMPK, p-AKT, and β-actin protein levels in liver tissues from different groups were measured by western blot. Representative of three experiments. b p-AMPK, p-AKT, and β-actin protein levels in KCs isolated from different groups were measured by western blot. Representative of three experiments. c KCs isolated from APAP and STZ + APAP mice were treated with PI3K antagonist LY294002 (20 µM) for 6 h in vitro. Intracellular p-AKT, and β-actin protein levels were measured by western blot. Representative of three experiments. d ROS production was detected by CarboxyH2DFFDA in KCs (original magnification ×200). Positive green fluorescent-labeled cells were counted blindly in 10 HPF/section (×200). e Quantification of ROS-producing KCs (green) per high power field (×200) (n = 3/group). Both CON and STZ mice were pretreated with AMPK activator (AICAR) (AICAR, 100 mg/kg, i.p.) once a day for 7 days prior to TAA administration or saline in vivo prior to APAP treatment. f Intracellular p-AMPK, p-AKT, and β-actin protein levels were measured by western blot. Representative of three experiments. g ROS production was detected by CarboxyH2DFFDA in KCs (original magnification ×200). Positive green fluorescent-labeled cells were counted blindly in 10 HPF/section (×200). h Quantification of ROS-producing KCs (green) per high power field (×200) (n = 3 /group) (mean ± SD, *p < 0.05)