Ginseng Berry Extract Prevents Atherogenesis via Anti-Inflammatory Action by Upregulating Phase II Gene Expression
Figure 6
KGBE inhibits intracellular ROS levels and NF-κB activation in LPS-stimulated ApoE−/− macrophages. ApoE−/− macrophages were pretreated with 100 μg/mL KGBE in the presence or absence of 20 μM SnPP and 20 μM BSO for 30 min and stimulated with LPS for 1 h. (a) Intracellular ROS accumulation was determined using DCFH2-DA. (b, c) Nuclear translocation of NF-κB was determined in whole cell mounts and nuclear and cytosolic fractions using confocal microscopy and Western blotting. (d) Nuclear NF-κB-DNA-binding activity was determined in the presence or absence of cold probe (CP) or antibody for NF-κB p65 (Ab) by EMSA. (e) Macrophages were transfected with piNOS-Luc or pNF-κB-Luc using a lipofectamine method and stimulated with LPS for 16 h following pretreatment with KGBE alone or in combination with SnPP and BSO for 30 min. Luciferase activity was measured in cell lysates by a luminometer. (f) Macrophages were pretreated with KGBE alone or in combination with SnPP and BSO in the presence or absence of 5 μM MG132 for 30 min, followed by stimulation with LPS for 30 min. IκB phosphorylation and degradation and IKK phosphorylation were determined by Western blotting. Data shown in (a, e) are the mean ± S.D. (). * and **.
