Figure 9
A1AT and LPS/A1AT induce PPAR-γ expression; effect of GW9662 on A1AT-LA ability to inhibit LPS-induced IL-1β expression Human neutrophils (5 × 106) were incubated for 5 h in either medium alone, medium containing A1AT (Prolastin, 1 mg/mL) or LPS (20 ng/mL) in the absence or presence of A1AT-0 (1 mg/mL) or A1AT-LA (1 mg/mL). Expression of PPAR-γ (A–D) was analyzed by real-time qPCR as described in Materials and Methods. GAPDH and HPRT were used as housekeeping genes. Box plots represent data from 3 individual donors; n = number of replicates for each experiment. P value indicates significant differences compared with the values seen in control cells (A and B) or LPS (C and D). (E) Cells were pretreated for 30 min with 10 µM GW9662, an irreversible PPAR-γ antagonist, prior to addition of LPS or LPS/A1ATs. Data are expressed as percentage of IL-1β gene expression in cells activated with LPS (100%) and show means (SD) of 3 independent donors; n = number of replicates for each experiment. P < 0.001 represents significant difference compared with the values seen in LPS-activated cells.