Coagulation factor Xa synergistically interacts with serotonin in inducing vascular smooth muscle cell proliferation

Abstract

Introduction

Initial events following vascular interventions include activation of platelets and coagulation cascade. Activated platelets release several vasoactive mediators including serotonin. Activation of coagulation cascade results in conversion of inactive zymogens such as factor X to its active form (factor Xa). So this study designed to examine the effect of factor Xa on rabbit vascular smooth muscle cell (VSMC) proliferation and its interaction with serotonin.

Methods

Growth-arrested VSMCs were incubated in a serum-free medium with different concentrations of factor Xa with or without serotonin. VSMC proliferation was examined by increase in incorporation of [³H]thymidine into DNA and by increase in cell number.

Results

Factor Xa and serotonin stimulated DNA synthesis in a dose-dependent manner. Factor Xa had a maximal effect at 100 nM (1180±110%) and serotonin at 50 μM (345±21%). When added together, at nonmitogenic concentrations, factor Xa (0.1 nM) and serotonin (1 μM) synergistically induced DNA synthesis (312±12%). These increases in DNA synthesis were paralleled by an increase in cell number. Serine protease inhibitors, active site blockers and platelet-derived growth factor receptor tyrosine kinase inhibitor blocked the mitogenic effect of factor Xa and its interaction with serotonin. Similarly, serotonin type 2 receptor inhibitor and Gi-protein-coupled receptor inhibitor inhibited the mitogenic effect of serotonin and its interaction with factor Xa. When used in combination, they blocked the interaction between factor Xa and serotonin.

Conclusion

Coagulation factor Xa and serotonin are mitogenic to VSMCs and also function as amplification factors to each other, suggesting that inhibition of neointimal proliferation after vascular injury may require the combined use of multiple growth factor inhibitors to simultaneously block several critical cellular activation pathways.