Systems pharmacology approach reveals protective mechanisms of Jian-Pi Qing-Chang decoction on ulcerative colitis

Figure 6 Jian-Pi Qing-Chang regulates tight junctions via the NF-κB/HIF-1α signalling pathway in Caco2 cells. A: The cell co-culture assay. Bone marrow-derived macrophages (BMDMs) were co-cultured in the upper chamber of a trans-well plate, with Caco2 cells placed in the lower chamber on day 5. Mature BMDMs were treated with TNF-α (20 ng/ml) in the presence or absence of either Jian-Pi Qing-Chang (100 μg/mL) or RAP (2 μM) for 24 h, or pre-treated with pyrrolidinedithiocarbamate (50 μM) for 30 min; B: The relative mRNA expression in Caco2 cells was measured by PCR; C-F: The relative protein expression in Caco2 cells was measured by Western blot; G and H: Double immunofluorescence staining for HIF-1α/Tublin and NF-κB/Claudin-1 in Caco2 cells. Data are presented as the mean ± SD. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. BMDMs: Bone marrow-derived macrophages; JPQC: Jian-Pi Qing-Chang; PDTC: Pyrrolidinedithiocarbamate; RAP: Rapamycin.