FIGURE 7.

Koumine (KM) inhibits ROS generation triggered by LPS and ATP. (A,B) BMDMs were pretreated with various concentrations of KM for 1 h and then incubated with LPS (300 ng/ml) for 3 h and ATP (5 mM) for 1 h. Intracellular ROS were labeled with the DCFH-DA probe and examined by fluorescence microscopy. Scale bars, 25 μm. Mean ± SD of three independent experiments are shown. #p < 0.05 vs. vehicle group, *p < 0.05 vs. LPS plus ATP group. (C,D) BMDMs were pretreated with KM (200 μM) for 1 h and then incubated with LPS (300 ng/ml) for 3 h and ATP (5 mM) for 1 h. H₂O₂ (10 mM) was added to cells at the same time as KM. Intracellular ROS were labeled with a DCFH-DA probe and examined by flow cytometry. #p < 0.05 vs. vehicle group, *p < 0.05 vs. LPS plus ATP group, $p < 0.05 vs. KM plus LPS and ATP group. (E,F) The protein levels of total AMPK and its phosphorylated forms (p-AMPK) were analyzed by western blotting. Mean ± SD of three independent experiments are shown. #p < 0.05 vs. vehicle group.