Coagulation factor XI induces Ca²⁺ response and accelerates cell migration in vascular smooth muscle cells via proteinase-activated receptor 1.

Activated coagulation factor XI (FXIa) is a serine proteinase that plays a key role in the intrinsic coagulation pathway. The analysis of FXI-knockout mice has indicated the contribution of FXI to the pathogenesis of atherosclerosis. However, the underlying mechanism remains unknown. We hypothesized that FXIa exerts vascular smooth muscle effects via proteinase-activated receptor 1 (PAR₁). Fura-2 fluorometry revealed that FXIa elicited intracellular Ca²⁺ signal in rat embryo aorta smooth muscle A7r5 cells. The influx of extracellular Ca²⁺ played a greater role in generating Ca²⁺ signal than the Ca²⁺ release from intracellular stores. The FXIa-induced Ca²⁺ signal was abolished by the pretreatment with atopaxar, an antagonist of PAR₁, or 4-amidinophenylmethanesulfonyl fluoride (p-APMSF), an inhibitor of proteinase, while it was also lost in embryonic fibroblasts derived from PAR₁^-/- mice. FXIa cleaved the recombinant protein containing the extracellular region of PAR₁ at the same site (R45/S46) as that of thrombin, a canonical PAR₁ agonist. The FXIa-induced Ca²⁺ influx was inhibited by diltiazem, an L-type Ca²⁺ channel blocker, and by siRNA targeted to Ca_V1.2. The FXIa-induced Ca²⁺ influx was also inhibited by GF109203X and rottlerin, inhibitors of protein kinase C. In a wound healing assay, FXIa increased the rate of cell migration by 2.46-fold of control, which was partly inhibited by atopaxar or diltiazem. In conclusion, FXIa mainly elicits the Ca²⁺ signal via the PAR₁/Ca_V1.2-mediated Ca²⁺ influx and accelerates the migration in vascular smooth muscle cells. The present study provides the first evidence that FXIa exerts a direct cellular effect on vascular smooth muscle.

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