Downregulation of Aquaporin 3 Mediated the Laxative Effect in the Rat Colon by a Purified Resin Glycoside Fraction from Pharbitis Semen

Figure 6

RFP activated NF-κB pathway and their downstream proteins in HT-29 cells. (a, b) Indicated concentrations of RFP were applied to treat HT-29 cells. (a) Western blot analysed the transposition of p-p65 in the nuclear and cytoplasm COX-2. PCNA and GAPDH were used as an equal loading control. P < 0.001, ns vs control group. (b) PGE₂ concentration was determined with ELISA kit, and the control cells presented as 100%. The data represented the means ± SDs for three experiments, P < 0.001, ns vs control group. (c, d) Cells were pretreated with NF-κB inhibitor BAY 11-7082 (BAY) (10 μM) or COX-2 inhibitor indomethacin (20 μM) for 1 h and further treated with RFP (4 μg/ml). (c) The expressions of p-p65, COX-2 and AQP3 were detected by western blotting. P < 0.01, P < 0.001, ns vs control group. (d) PGE₂ concentration was measured with the same method above. The data represented the means ± SDs for three experiments, P < 0.001, ns vs control group.

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