The Dietary Flavonoid Kaempferol Mediates Anti-Inflammatory Responses via the Src, Syk, IRAK1, and IRAK4 Molecular Targets

Figure 2

The effects of KF on iNOS, COX-2, and TNF-α gene expression and transcriptional regulation in LPS-treated RAW264.7 cells. (a) RAW264.7 cells (5 × 10⁶ cells/mL) were incubated with LPS (1 μg/mL) in the presence or absence of KF for 6 h. iNOS, COX-2, and TNF-α mRNA levels were determined using real-time PCR. (b) RAW264.7 cells (5 × 10⁶ cells/mL) were incubated with LPS (1 μg/mL) in the presence or absence of KF for the indicated times. After preparing the nuclear fractions, the translocated levels of total transcription factors (p65, p50, c-Fos, and c-Jun) were identified using immunoblot analyses. (c) HEK293 cells cotransfected with the NF-κB-Luc or AP-1-Luc (1 μg/mL each) and β-gal (as a transfection control) plasmid constructs were treated with KF in the presence or absence of adaptor molecule (MyD88 or TRIF) for 12 h. Luciferase activity was determined using luminometry. Relative intensity was calculated using total levels by the DNR Bio-Imaging System. All of the data are expressed as the mean ± SD of experiments that were performed with six or three (b) samples. and compared to the control group.

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