Effects of Tanshinone IIA on procoagulant activity of human ECV304 cell line induced by NB4 cells

Objective To investigate the effects of Tanshinone ⅡA (TanⅡA) on procoagulant activity (PCA) of human ECV304 cells induced by acute promyelocytic leukemia cell line NB4 cells. Methods ECV304 monolayers were respectively incubated for different hours at 37 ℃ in the conditioned media (CM) of NB4 cells treated with 0.5 μg/mL Tan ⅡA (Tan ⅡA -NB4-CM), 0.3 μg/mL all-trans retinoidic acid (ATRA)(ATRA-NB4-CM), DMSO(DMSO-NB4-CM) or the RPMI1640 medium. ECV304 lysates were tested for PCA using the one-stage clotting assay as well as for tissue factor activity (TF: Act) using the chromogenic substrate assay; ECV304 cell monolayers were incubated for different hours at 37 ℃ in a medium system including 0.5 μg/mL TanⅡA and TanⅡA -NB4-CM, and the ECV304 cell lysates were tested for PCA in the same way as above. Also they were controlled by 0.3 μg/mL ATRA, DMSO or RPMI1640 medium. Results ①The conditioned mediums from 0.5 μg/mL TanⅡA that treated NB4 cells for 24, 72 and 120 hours respectively could elevate PCA of ECV cells, and this capability developed with the time of reaction. ATRA did the same as TanⅡA (P0.05). ② 0.5 μg/mL TanⅡA down-regulated the PCA of ECV304 cells induced by TanⅡA -NB4-CM, and the inhibitory effects increased with time, reaching the highest at 120 hours. ③ TanⅡA 120 h-NB4-CM up-regulated TF:Act of ECV304 cells, and the effect increased with time. ④ 0.5 μg/mL TanⅡA down-regulated PCA and TF:Act of ECV304 cells induced by TanⅡA -NB4-CM, and the inhibitory effect increased with time; simultaneously, the test was controlled with 0.3 μg/mL ATRA, the effects on PCA and TF:Act were not significantly different (P0.05). Conclusion TanⅡA -NB4-CM can increase the levels of PCA and TF:Act of ECV304 cells through some unidentified factor; however, TanⅡA can obviously decrease the PCA and TF:Act levels of ECV304 cells induced by TanⅡA -NB4-CM.