Fig. 4.

TGR5 activation inhibited TLR4-mediated pro-inflammatory responses in macrophages. BMDMs were isolated from TGR5-proficient and TGR5-deficient mice, cultured and differentiated for 7 days, and then stimulated with LPS for 24 h. INT-777 was added 1 h before LPS stimulation. (A–C) TNF-α, IL-6 and IL-10 mRNA levels were measured with qRT-PCR (n = 4), and (D–F) serum cytokine levels were detected with ELISA (n = 4 for each group). (G) TLR4, p-NF-κB p65, p-Akt, p-Gsk3β and β-actin were detected by western blot (n = 3). Statistical analysis was performed by Student’s t-test. Each column represented the mean ± SD. *P < 0.05, **P < 0.01.