OroA inhibition of LPS-induced NF-κB activation in lung tissue of the rats.

<p>LPS treatment (10 mg/kg, iv) significantly decreased cytosolic NF-κBp65 expression in lung tissue (Lane 2 in Fig. 6A) comparing to that by Sal (Saline) followed by Solv (Solvent) treatment (Lane 1 in Fig. 6A). The reduction examined (E) 24 hrs after LPS challenge (Fig. 6A) was significantly reversed to similar extent by treatment with OroA (15 mg/kg, iv) administered 1 hr (Lane 3 in Fig. 6A) or 6 hrs (Lane 5 in Fig. 6A) after LPS. Bar graphs summarized the relative density (%) of cytosolic NF-κB protein that was normalized to actin (Fig. 6A). In parallel, LPS treatment (10 mg/kg, iv) significantly increased nuclear p65 expression (Lanes 2 and 4 in Fig. 6B) comparing to that by Sal followed by Solv treatment (Lane 1 in Fig. 6B). The increase examined 24 hrs after LPS challenge was significantly reversed to similar extent by treatment with OroA (15 mg/kg, iv) administered 1 hr (Lane 3 in Fig. 6B) or 6 hrs (Lane 5 in Fig. 6B) after LPS challenge. Bar graphs summarized the relative density (%) of nuclear NF-κB protein that was normalized to H2A. Solvent (Solv, normal saline plus Tween 80 at 9∶1 ratio); hr/Posttreat (post-treatment hour after LPS treatment); E/hr (examination hour after LPS challenge). Data are means ± SEM. *P<0.05 indicates significant difference from the normal control (Sal followed by Solv, Lane 1 in panels A and B). #P<0.05 indicates significant difference from the respective LPS treated group (LPS followed by Solv) (Lanes 2 and 4 in Figs. 6A and 6B) when examined 24 hrs after LPS challenge. The number in each column represents the number of rats examined. Fig. 6Ci indicates effects of LPS (10 mg/kg, iv) administered 6 hrs after LPS challenge on phosphorylated p65 cells in red (indicated by yellow arrows in panels Ci and Cii) which were examined 24 hrs (E/24h) after LPS challenge. Phosphorylated NF-κB p65 cells were significantly reduced by OroA (15 mg/kg, iv) administered 6 hrs after LPS challenge (panel Cii). Phosphorylated NF-κBp65 cells were calculated from randomly selected 4 fields with a total of 200 cells. Insets are enlarged areas indicated by white arrowheads denoting phosphorylated NF-κBp65 translocated into the nuclei. The nucleus in blue color was stained with DAPI. Data are means±SEM. #P<0.05 indicates significantly different from the LPS plus Solv-treated group. Scale bar, 20 µm. The number in each column represents the number of rats examined.