Figure 1.

ASMase inhibitor treatment attenuates LPS-induced TF procoagulant activity and thrombin generation in vivo. Wild-type (C57BL/6J) mice were injected with saline (Con) or ASMase inhibitor desipramine (Des) or imipramine (Imi) (10 mg/kg body weight in 100 μL of saline) daily intraperitoneally for 7 days. After 7 days, mice were challenged with saline or LPS (E coli O111:B4, 5 mg/kg) by injecting them intraperitoneally in 100 µL volume. In a group of mice, 4 hours following LPS challenge, Bz-ATP (50 mg/kg in 100 µL) was administered. Fifteen minutes following ATP administration, animals were euthanized, and blood was drawn into citrate anticoagulant by cardiac puncture. Whole blood was loaded on a Ficoll gradient and centrifuged at 400g for 40 minutes to separate the plasma and PBMCs. PBMCs were washed and used to measure TF activity, and the plasma was processed for isolation of MVs or measuring thrombin generation. (A) TF activity of PBMCs. (B) TF activity associated with MVs. (C) Levels of TAT in plasma. Data are mean ± SEM (n = 6 mice per group). Differences between the group mean in all 3 panels, as determined by 1-way ANOVA, were highly statistically significant (P < .0001). *P < .05, **P < .01, ***P < .001, Tukey’s post hoc multiple comparisons.