Effects Of Tanshinone â…¡A On NB4cell-induced Procoagulant Activity In HUVECs

ObjectiveTo investigate the effect of tanshinoneⅡA on procoagulant activity(PCA) of human umbilical vein endothelial cells (HUVECs) induced by acute promyelocytic leukemia (APL) cell line NB4 cells.Material and MethodsHUVECs monolayers were respectively incubated for 6,12,24 hours with different tanshinoneⅡA conditioned medias(TanⅡA-NB4-CM), HUVECs lysates was obtained by freezing and thrawing repeatedly for 3times,and then their PCA was tested using the one-stage clotting assay.Then HUVECs monolayers were respectively incubated for 6,12,24,72 hours with TanⅡA-NB4-120h-CM and different concentration of TanⅡA, HUVECs lysates was obtained and the PCA was tested using the one-stage clotting assay,and TF:act was tested using the chromogenic substrate assay.All the above were controlled by 0.3μg/ml ATRA,0.01%DMSO and RPMI 1640.Resultsl.TanⅡA-(72h,120h)-NB4-conditioned media could elevat PCA of HUVECs,with the most obvious effect of 120h-NB4-CM,at 6th hour. PCA of HUVECs treated with 1.0μg/ml TanⅡA or 0.3μg/ml ATRA were no difference.2. The NB4-CM induced PCA of HUVECs could be effectively reduced by TanⅡA at the concentration of 5.0μg/ml,which had no difference with what was induced by ATRA at the concentration of 0.3μg/ml.The concentration of TanⅡA below 5.0μg/ml had no effect to reduce its NB4-CM induced PCA of HUVECs,which was differernt from that of 0.3μg/mlATRA.3.Both TanⅡA 120h-NB4-CM and ATRA 120h-NB4-CM could elevate the TF:act of HUVECs when incubated with them for 6 hours,while the TanⅡA 120 h-NB4-CM induced PCA remained to be at the same level at 12th hour as that of 6th hour, fell to base line at 24th hour, and the ATRA 120 h-NB4-CM induced TF:act stepped down with time after reach the peak at 6th hour.4. 1.0μg/mlTanⅡA could not reduce the TF:act of HUVECs,and 0.3ug/ml ATRA could obviously reduce it at 6th hour.Conclusion1.TanⅡA 120h-NB4-CM could increase the level of PCA and TF:act of HUVECs. PCA of HUVECs treated with TanⅡA and ATRA 120h-NB4-CM was no difference.2.PCA of HUVECs induced by TanⅡA 120h-NB4-CM could be decreased by TanⅡA at concentration of 5.0μg/ml,which was the same as that did by 0.3μg/ml ATRA. While,TF activity of HUVECs induced by NB4-CM could be decreased by ATRA at concentration of 0.3μg/ml,but,it was not decreased by TanⅡA at concentration of 1.0μg/ml...