title: Factor Xa in mouse fibroblasts may induce fibrosis more than thrombin.
abstract: Coagulation factors are known to play a role in wound healing by stimulating fibroblasts and might be associated with tissue fibrosis, however, only limited data exist. Protease-activated receptor 1 (PAR1), activated by thrombin or factor (F) Xa, and PAR2, activated by FXa, have recently been reported to play roles not only in the coagulation system, but also in cardiac fibrosis. Furthermore, a previous report found that FX deficiency in mice led to the development of cardiac fibrosis. Therefore, in the present study, we evaluated cellular biological function under conditions of overexpressed thrombin and FXa in fibroblasts.Cell migration and proliferation with FXa (1U/mL) and thrombin (1U/mL) stimulation were evaluated. Cells incubated without FXa or thrombin were used as control. H2O2 and TGF-β1 production were measured using ELISA. Signal pathways were evaluated using a signal pathway reporter assay.Cell migration and proliferation were increased in FXa-stimulated cells (4.1-fold increase for migration, 1.3-fold for proliferation compared with control, respectively) and thrombin (4.1-fold increase for migration, 1.3-fold for proliferation as compared to control, respectively). H2O2 production was higher in FXa-stimulated cells compared to thrombin (1.3-fold increase) and control cells (1.4-fold increased). TGF-β1 production was up-regulated after FXa addition (12.6-fold increase compared with thrombin, 1.8-fold increase compared with control, respectively). In FXa-stimulated cells, AP-1 and NF-kB were increased compared to control (P < 0.05).These data suggest that FXa and thrombin play important roles in the fibrotic process that could also lead to cardiac fibrosis, and that at least some of these signalings are more accelerated with FXa compared to thrombin.
authors.affiliation: Kitasato University Graduate School of Medical Sciences.
authors.forename: Lisa
authors.initials: L
authors.lastname: Kitasato
authors.forename: Minako
authors.initials: M
authors.lastname: Yamaoka-Tojo
authors.forename: Takehiro
authors.initials: T
authors.lastname: Hashikata
authors.forename: Sayaka
authors.initials: S
authors.lastname: Ishii
authors.forename: Ryo
authors.initials: R
authors.lastname: Kameda
authors.forename: Takao
authors.initials: T
authors.lastname: Shimohama
authors.forename: Taiki
authors.initials: T
authors.lastname: Tojo
authors.forename: Junya
authors.initials: J
authors.lastname: Ako
mesh_terms: D000818:Animals; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D004797:Enzyme-Linked Immunosorbent Assay; D015951:Factor Xa / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005355:Fibrosis; D006490:Hemostatics / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D015398:Signal Transduction; D013917:Thrombin / Q000494:pharmacology*; D018808:Transcription Factor AP-1 / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D006490:Hemostatics; D016328:NF-kappa B; D018808:Transcription Factor AP-1; D053773:Transforming Growth Factor beta1; D006861:Hydrogen Peroxide; D013917:Thrombin; D015951:Factor Xa