Diosgenin inhibits tumor necrosis factor-induced tissue factor activity and expression in THP-1 cells via down-regulation of the NF-κB, Akt, and MAPK signaling pathways

Abstract

AimTo investigate whether diosgenin could modulate tissue factor (TF) procoagulation activity, expression, and related signal transduction pathways. MethodsHuman THP-1 monocytic cells were exposed to tumor necrosis factor-α (TNF-α, 10 ng·mL−1) with or without diosgenin (0.01, 0.1, and 1 μmol·L−1) for 2 h or 5 h to induce TF procoagulant activity and expression, which were determined by the simplified chromogenic assay, reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR, and Western blotting assays. In addition, the activation of the NF-κB, Akt, and MAPK signaling pathways were also measured by Western blotting. ResultsDiosgenin significantly inhibited TNF-α-induced TF procoagulant activity at concentrations of 0.01 to 1 μmol·L−1 with IC50 of 0.25 μmol·L−1. It also reduced protein expression and mRNA accumulation of TF dose-dependently in activated THP-1 cells. TNF-α stimulated significantly phosphorylation on Ser536 of NF-κB/p65, Ser473 of Akt at 5–15 min, and activations of IKK-β and ERK at 15–30 min. Diosgenin (1 μmol·L−1) could inhibit the phosphorylation of NF-κB/p65, IKK-β, Akt, ERK, and JNK, but had no remarkable effects on IκB and p38 phosphorylation in THP-1 cells. ConclusionDiosgenin inhibits TNF-α-induced TF activity and expression in monocytes, partly due to its down-regulation of the phosphorylation of NF-κB/p65, IKK-β, Akt, ERK, and JNK.