摘要
目的研究丹参酮A(TAN A)对急性早幼粒细胞性白血病(APL)细胞株NB4诱导的人ECV304细胞株促凝活性的影响。方法1分别用0.5ΜG/ML TAN A、0.3ΜG/ML ATRA、0.1G/L DMSO、RPMI1640处理培养NB4细胞制成条件培养基TAN A-NB4-CM,ATRA-NB4-CM、DMSO-NB4-CM以及RPMI1640-NB4-CM,再分别与ECV304细胞在37℃共同孵育0、4、8、12H,利用复钙时间测定及改良发色底物法,分别测定ECV304细胞裂解液的肿瘤促凝活性(PCA)和组织因子活力(TF:ACT)。2ECV304细胞分别与0.5ΜG/ML TAN A及TAN A-NB4-CM在37℃共同孵育4、12、24、72、120H,用上述同样方法测定ECV304细胞裂解液的PCA和TF:ACT。结果10.5ΜG/ML TAN A处理NB4细胞24、72、120H的条件培养基能使ECV304细胞PCA增强,ATRA具有相似作用(P>0.05)。20.5ΜG/ML的TAN A对0.5ΜG/ML TAN A作用NB4120H的条件培养基(TAN A-120H-NB4-CM)促ECV304细胞PCA有抑制作用,其作用强度呈时间依赖性,120H达到高峰,再增加TAN A浓度,作用强度不增加;与0.3ΜG/ML ATRA作用相似。3TAN A-120H-NB4-CM能够增加ECV304细胞TF:ACT,并随作用时间增加而增强,TAN A与ATRA作用相似(P>0.05)。40.5ΜG/ML TAN A能够抑制TAN A-120H-NB4-CM对ECV304细胞的TF:ACT,并随作用时间增加而增强,ATRA具有相似作用(P>0.05)。结论TAN A在诱导NB4细胞分化凋亡的同时,可能通过某种因子增强NB4细胞对ECV304细胞促凝活性和组织因子活力;TAN A能够有效阻止NB4细胞增强ECV304细胞的促凝活性和组织因子活力,起到保护细胞的作用。
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 [A（TanⅡA-NB4-CM）, 0. 3μg/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 onestage 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. 5μg/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 0.3μg/mL ATRA, DMSO or RPMI1640 medium. Results ①The conditioned mediums from 0. 5μg/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 （P〈0. 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 TanⅡA120 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 0.3μg/mL ATRA, the effects on PCA and TF：Act were not significantly different （P〉0. 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 Tan ⅡA-NB4-CM.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2006年第1期55-59,共5页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(批准号30470748)资助
