Therapeutic Effects of Fermented Flax Seed Oil on NC/Nga Mice with Atopic Dermatitis-Like Skin Lesions

Figure 3

Downregulation of inflammation. (a1) Cells were pretreated with different concentrations of FFSO for 30 minutes and then stimulated with LPS (1 microg/mL) for 12 hours. Ratio of TNF-α to GAPDH expression in cells was determined by real-time PCR, as described in Section 2. (a2) Cells were pretreated with the indicated concentrations of FFSO for 2 hours and then stimulated with LPS (1 microg/mL) for 1 hour, after which nuclear extracts were prepared. Detection of NF-κB-binding activities was performed as described in Section 2. This experiment was repeated two times with similar results. (a3) Ratio of COX-2 to GAPDH expression on cells same as (a1). (a4) Cells were pretreated with different concentrations of FFSO for 30 minutes and then stimulated with LPS (1 microg/mL) for another 24 hours. Expression of iNOS and COX-2 proteins was detected by Western blotting using specific anti-iNOS and anti-COX-2 antibodies. β-Actin protein was used as an internal control. Expression of TNF-α, NF-kB p65, iNOS, and COX-2 (arrow indicates dark brown) was induced by D. farinae in the AE group. These positive reactions in the AT group were remarkably decreased compared with those in the AE group (immunohistochemistry; Bar size, 50 μm). Abbreviations are the same as in Figure 1. compared with AE; compared with AE; compared with Ctrl.