Lumbrokinase, a Fibrinolytic Enzyme, Prevents Intra-Abdominal Adhesion by Inhibiting the Migrative and Adhesive Activities of Fibroblast via Attenuation of the AP-1/ICAM-1 Signaling Pathway

Figure 4

Lumbrokinase inhibits the AP-1/ICAM-1 signaling pathway. (a) IMR-90 cells () were seeded into a 24-well culture plate. One day after incubation, the cells were treated with lumbrokinase (2,000 U/mL) for two days. The cells were then transfected with reporter plasmids containing the promoter of AP-1, β-catenin (OT), ERK (ARE), and NF-κB. The cells were harvested two days after transfection, and the transcriptional activity was measured using a dual-luciferase reporter assay kit. and . (b) 5 × 10⁵ IMR-90 cells were seeded in 6-well plates, then treated with the indicated concentration of lumbrokinase after 24 h. After 48 h of incubation, the cells were harvested for western blot analysis. (c) IMR-90 cells were incubated with or without lumbrokinase (2,000 U/ml) in the presence and absence of phorbol 12-myristate 13-acetate (PMA), an AP-1 enhancer, for 48 h. Then, the cells were harvested and subjected to western blotting. The graph was plotted based on the band densities measured using the ImageJ software. and . (d) IMR-90 cells were plated and treated with lumbrokinase (2,000 U/mL) and T-5224 (an AP-1 inhibitor) 24 h later. The cells were harvested after 48 h of incubation in their respective, after which western blotting was performed. The graph was plotted based on the band densities measured using the ImageJ software. and .

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