Effect of Tetrandrine on LPS-induced NF-κB activation in isolated pancreatic acinar cells of rat

Figure 1 Protective effect of Tet on LPS-induced cell damage in pancreatic acinar cells. Pancreatic acinar cells were treated with LPS 10 mg/L, Tet 50 μmol/L or Tet 100 μmol/L for 1, 4, and 10 h. Results are expressed as percentage of control values (without LPS treatment) at 0 h time-point (n = 3). Mean ± SD. ^aP < 0.05, ^bP < 0.01 vs LPS group at the same time-point. ^cP < 0.05, ^dP < 0.01 vs control group at the same time-point.

Figure 2 Tet prevented LPS-induced nuclear translocation of p65 in isolated pancreatic acinar cells. P65 was visualized by indirect immunofluorescence staining using rabbit anti-p65 polyclonal antibodies (1:100) which only recognized NF-κB p65. Goat anti-rabbit antibodies (1:100) conjugated to FITC was performed, and visualized under a confocal microscope. Nuclear p65 was observed at 1 h (C), 4 h (D) after treatment with LPS (10 mg/L) but not in saline-treated controls at 1 h (B), nor in negative control (A). Nuclear localization of p65 and the fluorescence in cytoplasm were markedly reduced by Tet 100 μmol/L at 1 (E), 4 h (F), compared with that of LPS group at the same time (Original magnification × 400).

Figure 3 A: Effect of LPS on NF-κB activity in pancreatic acinar cell. Lane 1: control; Lane 2: treated with LPS 10 mg/L for 1 h; Lane 3: treated with LPS 10 mg/L for 1 h + cold NF-κB probe; Lane 4: treated with LPS 10 mg/L for 1 h + labeled AP₂ probe; B: The time course of LPS-induced NF-κB activity in pancreatic acinar cell. NF-κB activity in pancreatic acinar cell stimulated by LPS 10 mg/L for indicated periods of time. Lane 1: 0 min; Lane 2: 30 min; Lane 3: 1 h; Lane 4: 4 h; Lane 5: 10 h; C: Effect of Tet on LPS-induced NF-κB activity in pancreatic acinar cell. Lane 1: control; Lane 2: treated with Tet 100 μmol/L for 1 h; Lane 3: treated with LPS 10 mg/L for 1 h; Lane 4: treated with LPS 10 mg/L + Tet 100 μmol/L for 1 h; Lane 5: treated with LPS 10 mg/L for 4 h; Lane 6: treated with LPS 10 mg/L + Tet 100 μmol/L for 4 h.