CCK reduces LPS-induced NF-κB activation and p65 translocation to nucleus and also delays IκBα degradation by peritoneal macrophages. The macrophage culture was pretreated with CCK (10−10 or 10−6 M) for 30 min before the addition of LPS (1 μg/mL) to the cell culture medium (time zero). At the indicated time-points, nuclear proteins were extracted for the quantification of p65 translocation by using ELISA kit (a). The nuclear p65 concentration was presented as the percentage relative to the control (saline) group. Cytosolic proteins were also extracted for the determination of p65 content and IκBα degradation by western blot. Representative immunoblots with densitometry values were showed for p65/β-actin ratio (b) and IκBα/α-tubulin ratio (c). The β-actin and α-tubulin levels were used as internal controls. Data is a representative of three independent experiments with similar results. Values are expressed as means ± S.E.M. 4–10 samples per group. * versus saline and # versus LPS group.
