Astaxanthin Inhibits Interleukin-6 Expression in Cerulein/Resistin-Stimulated Pancreatic Acinar Cells

Figure 6

NAC, ML171, or BAPTA-AM inhibits cerulein/resistin-induced increases in ROS, NADPH oxidase activity, and nuclear translocation of NF-κB p65 in AR42J cells. The cells were pretreated with NAC (1 mM) or ML171 (2 μM) or BAPTA-AM (5 μM) for 1 h and then stimulated with cerulein/resistin for 45 min (for ROS levels, ROS fluorescence imaging, and NADPH oxidase activity (a–c)), and for 1 h (for nuclear level of NF-κB p65 by confocal images (d)). (a) Intracellular ROS levels were measured by dichlorofluorescein diacetate (DCF-DA) fluorescence. (b) Representative images of ROS-induced fluorescence response of DCF-DA. (c) NADPH oxidase activity in the membrane fraction was measured by the lucigenin assay. Data are expressed as the of three different experiments. ROS level and NADPH oxidase activity in none (untreated cells) were set as 100%. vs. none (untreated cells); ⁺ vs. cerulein (cells treated with cerulein alone). (d) The immunoreactive NF-κB p65 was visualized using a rhodamine-conjugated mouse anti-rabbit IgG antibody (red) with DAPI counterstaining (blue) of the same field.

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