Differential Regulation of NF-κB and Nrf2 by Bojungikki-Tang Is Associated with Suppressing Lung Inflammation

Figure 3

BT activates the transcriptional activity of Nrf2. (a) RAW 264.7 cells were treated with varying amounts of BT for 16 h. As for Nrf2 control, cells were similarly treated with 5 μM of sulforaphane (SFN), a potent Nrf2 activator. Nuclear proteins were isolated from these cells and analyzed by immunoblotting of Nrf2. The membrane was stripped and blotted with α-lamin A/C antibody for nuclear protein controls. A similar experiment was repeated at least three times. (b) RAW 264.7 cells were treated similarly to (a), and total RNA was extracted and analyzed by a semiquantitative RT-PCR for the expression of HO-1, GCLC, and NQO-1. (c) PCR bands in (b) were analyzed by ImageJ, and relative expressions of individual genes were shown in graphs after being normalized to GAPDH. (d) RAW 264.7 cells were treated with BT for 16 h and subsequently with TLR4-specific LPS (100 ng/ml) for 4 h. A semiquantitative RT-PCR was performed for the expression of HO-1, GCLC, and NQO-1. Resultant PCR bands were analyzed by ImageJ, and relative expressions of individual genes were shown in graphs after being normalized to GAPDH (e). Data represent the mean ± SEM of three independent measurements. was less than 0.05, compared to untreated controls.

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