Figure 5.

R17 inhibits lipid‐induced oxidative stress and ER stress. Panels (a–c), effects of R17 in HF‐fed mice as described for Figure 1. (a–b) Effects of R17 on the MDA level and SOD activity in the liver of mice. (c) Effects on UPR pathway proteins. Data shown are individual values with means ± SEM, N = 8 mice per group. *P < .05, significantly different from CH control mice; ^# P < .05, significantly different from HF control mice. Panels (d–g), HuH7 cells were incubated with OA (0.5 mM) or PA (0.5 mM) in the absence or presence of R17 (1 μM). (d–e) Effect on ROS level. ROS levels were determined by a ROS probe in flow cytometry. Representative images of DAPI (blue) for nucleus and ROS (green) were captured in confocal microscopy at 24 hr. (f) Effect on MDA level. (g) Effects on ER stress. The UPR proteins (PERK, elF‐2α, IRE‐1α, XBP‐1s, and CHOP) were immunoblotted with specific antibodies. Protein levels were quantified and normalized to actin; and relative fold increases were determined by comparison with the blank group. Panels (h–j), HuH7 cells were incubated with DTT (2 mM, ER stress inducer) in the absence or presence of R17 (0.4, 1, and 2.5 μM) for 2 hr. (h) Effects on UPR proteins; protein levels were quantified and normalized to actin; and relative fold increases were determined by comparison with the blank group. (i) Cell apoptosis. Determination by flow cytometry using an Annexin V‐FITC/PI kit; (j) cell viability. Data shown are individual values with means ± SEM, N = 5 independent experiments.*P < .05, significantly different from blank; ^# P < .05, significantly different from control (Ctrl)