Figure 3.

R17 inhibits lipid‐induced inflammation and macrophage accumulation. Panels (a–e), effects of R17 in HF‐fed mice as described for Figure 1. (a) Plasma levels of enzymes from the liver. (b) CD68 (macrophage marker as illustrated by arrows, top) and MAC387 (monocyte marker as illustrated by arrows, bottom) in the liver and labelled cell were quantified by Image J software as described. (c) mRNA levels of inflammatory cytokines in the liver. (d–e) Plasma levels of inflammatory cytokines determined by ELISA. 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 (f–h), HuH7 cells were incubated with OA (0.5 mM) or PA (0.5 mM) for 24 hr with or without R17 (1 μM) treatment. (f–g) Secretion level of TNF‐α and IL‐6. Culture medium was collected for the determination of cytokine secretion by an ELISA assay. (h) Protein levels of TNF‐α, IL‐6, and NF‐κB in cells were determined by immunoblotting with specific antibodies. Protein levels were quantified and normalized to actin; and relative fold increases were determined by comparison with the blank group. (i–j) Additive effects with the NF‐κB inhibitor SC75741 on TNF‐α, IL‐6, and NF‐κB. SC75741 (10 μM) was co‐incubated with R17 (1 μM) for 24 hr, cells were subjected to mRNA level (RT‐PCR) and protein level (immunoblotting) determination. Protein levels were quantified and normalized to actin; and relative fold increases were determined by comparison with the Ctrl group. 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); ^@ P < .05, significantly different from R17‐ or SC75741‐treated cells