Sikyungbanha-Tang Suppressing Acute Lung Injury in Mice Is Related to the Activation of Nrf2 and TNFAIP3

Figure 5

Activation of TNFAIP3 and suppression of NF-κB by SKBHT. (a) RAW 264.7 cells were treated with indicated amounts of SKBHT for 16 h from which the expression of TNFAIP3 was analyzed by immunoblotting. The membrane of TNFAIP3 was stripped and reblotted for β-actin. (b) RAW 264.7 cells pre-treated as in (a) were treated with LPS (100 ng/ml) for 15 min or 30 min, from which nuclear proteins were extracted. p65 RelA of NF-κB in the nucleus was analyzed by immunoblotting. The membrane was stripped and reblotted for lamin B for nuclear protein controls. The band intensity of p65 RelA and lamin B was measured three times by Image J The relative levels of p65 (NF-κB) over lamin B are shown in columns. P was less than 0.001, compared with the LPS only (post-ANOVA comparison with Tukey’s post hoc test). (c) RAW 264.7 cells were treated with SKBHT for 16 h and subsequently with LPS for 4 h Total RNA was extracted and analyzed by qPCR for IL-6, IL-1β, and TNF-α. Data represent the mean ± SEM of three independent measurements. P was less than 0.05, compared with the LPS-treated (post-ANOVA comparison with Tukey’s post hoc test).

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