THP1 cells were pretreated with hBD3 with or without H89, a PKA inhibitor (10μM) and hBD3 (10ug/ml) together or hBD3 alone for 1hr before the addition of LPS(200ng/ml). (A, B ) IL-6 and TNF-alpha were measured by ELISA 12hrs after LPS treatment. (C) IFN-beta expression was analyzed after 2hrs of LPS stimulation byReal time PCR and normalized against GAPDH expression (Mean ± SD of at least two independent experiments). (D, E) THP1 cells were treated with hBD3 (10μg/ml)for the indicated time points and phosphorylation of PKA and CREB was analysed by Western blots. Significant differences comparing with or without addition of H89 inhibitor and hBD3 were assessed by nonparametric Mann-Whitney U test and are indicated by *P<.05, **P<.01.
