Figure 1. ET Impairs LPS-Induced sPLA2-IIA Expression, but Not IL-8, PGE2, and NF-κB Translocation.

(A) AMs were pretreated with ET (PA + EF) for 1 h before overnight stimulation with P. aeruginosa LPS (50 ng/ml). ET consisted of a fixed concentration of PA (1 μg/ml) and increasing concentrations of EF.

(B) AMs were incubated with 1 μg/ml of ET (PA 1 μg/ml + EF 1 μg/ml), and were then stimulated overnight with TNFα (500 ng/ml). The sPLA2-IIA secretion levels were then assessed, as detailed in the Materials and Methods.

(C–E) After 1-h pretreatment with 1 μg/ml of ET (PA 1 μg/ml + EF 1 μg/ml), followed by an overnight stimulation with LPS (50 ng/ml), sPLA2-IIA mRNA levels and IL-8 and PGE2 concentrations were measured in culture medium. (C) shows reverse-transcription PCR (RT-PCR) analysis of sPLA2-IIA mRNA expression of extracts from three independent experiments; (D) and (E) show IL-8 and PGE2 concentrations, respectively.

(F) AMs were incubated with ET for 1 h and then were stimulated with LPS. After a 2-h incubation, NF-κB translocation was measured by EMSA.

The data are the mean ± standard error of the mean (SEM), and are representative of four separate experiments. An asterisk (*) indicates p < 0.05.