Total Flavonoids from Radix Glycyrrhiza Exert Anti-Inflammatory and Antitumorigenic Effects by Inactivating iNOS Signaling Pathways

Figure 3

Effect of TFRG on NF-κB, MAPK activities, and microRNA-155 expression in LPS/IFNγ-stimulated RAW264.7 cells. (a, b) DNA binding activity of p50 and p65 proteins in nuclear extracts was assessed using NF-κBp50/p65 EZ-TFA transcription factor assay. Absorbance was measured at 450 nm in a microplate spectrophotometer. Results were normalized to absorbance/mg protein. (c) RAW264.7 cells were plated at a density of 1 × 10⁶ cells/well in 30-mm dish overnight. TFRG were added to cells for 1 h followed by 30-min stimulation of IFNγ (10 U/ml) plus LPS (100 ng/ml). (d) Effect of TFRG on expression of miRNA-155 in stimulated RAW264.7 cells. The cells were stimulated with LPS plus IFN-γ only or stimulated with different concentrations (20, 40, 60 μg/ml) of TFRG for 18 h. Total RNA was isolated and the expression of miRNA-155 was determined by qPCR. RNU6B was used here as an endogenous control. The data represent the mean ± SEM of triplicate experiments. and compared with the model group alone.

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