title: 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: OBJECTIVE
  To investigate whether diosgenin could modulate tissue factor (TF) procoagulation activity, expression, and related signal transduction pathways.
  
  
  METHODS
  Human 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.
  
  
  RESULTS
  Diosgenin 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.
  
  
  CONCLUSIONS
  Diosgenin 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.
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Hao-Peng
authors.initials: HP
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yue
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Wen-Wen
authors.initials: WW
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China. Electronic address: junpingkou@163.com.
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Kou
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Bo-Yang
authors.initials: BY
authors.lastname: Yu
mesh_terms: D004144:Diosgenin / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
chemical_list: D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D051057:Proto-Oncogene Proteins c-akt; D004144:Diosgenin
keywords: Cardiovascular diseases; Diosgenin; Monocytes; TNF-α; Tissue factor