Fig 5. P.g. LPS Up-regulates Expression of Inflammatory Mediators via NF-κB Signaling in Trophoblasts.

(A) HTR-8 trophoblasts were seeded (5x10⁵ cells/well) in 6-well culture plates and culture media were changed once before stimulation. Cells were treated by P.g.-LPS (1 μg/ml unless otherwise noted) and both culture medium and cells were collected. mRNA expressions of COX-2, IL-8 and TNF-α were analyzed from cell pellets. (B) HTR-8 cells were stimulated with P.g.-LPS, Li-P.g. (live-P.g.) or D-P.g. (dead-P.g.) and cell lysates were examined by immunoblotting analysis using p-p65, p65. Molecular weight is labeled to the right of each band. (C,D) HTR-8 cells were pretreated with or without CAPE for 4 hrs and then stimulated with or without P.g.-LPS, Li-P.g. (live-P.g.) or D-P.g. (dead-P.g.) for 24 hrs. mRNA expressions of COX-2, IL-8 and TNF-α (C); protein secretion of TNF-α were examined (D). TNF-α amount in negative controls were subtracted as basal levels when calculating the percentage of down-regulation. GAPDH or β-actin was used as internal control. MOI, multiplicity of infection. *p < 0.05, **p < 0.01. Experiments were performed at least three times with similar results.