论文部分内容阅读
目的:研究丹参酮ⅡA(TanⅡA)对急性早幼粒细胞白血病(APL)细胞株NB4细胞诱导的血管内皮细胞株(ECV304)促凝活性(PCA)的影响,并对其机制作初步探讨。方法:(1)分别用1.0μg/mL TanⅡA、0.3μg/mLATRA、0.01%DMSO、PRMI1640处理NB4细胞24、48和72h,取其上清液作为条件培养基(hNB4-CM)。将这些CM分别与ECV304细胞在37oC共同孵育0、4、8和12h,用反复冻融法制备ECV304细胞裂解液,采用一期凝血法测定其PCA;采用ELISA法测定条件培养基中的TNF-α。(2)ECV304细胞与1.0μg/mL TanⅡA及TanⅡA 72h-NB4-CM在37oC共同分别孵育6、12、24和48h,并以ATRA和DMSO分别作为阳性和阴性对照,用上述相同方法测定ECV304细胞裂解液的PCA。结果:(1)1.0μg/mL TanⅡA可以诱导NB4细胞分化,其作用NB4细胞的培养基有一定的升高ECV304细胞PCA的作用,该作用在孵育4h时达高峰,之后ECV304细胞PCA逐渐下降。与0.3μg/mL ATRA的作用无统计学差异(P>0.05)。(2)1.0μg/mL的TanⅡA对TanⅡA-72h-NB4-CM促ECV304细胞PCA有抑制作用,其强度随作用时间增加而增加,与1.0μmol/L ATRA比较,P>0.05。(3)TanⅡA作用NB4细胞的培养基中TNF-α浓度,在作用前7h内随作用时间增加而增加,与0.3μg/mL ATRA比较无差异(P>0.05)。结论:TanⅡA能诱导NB4细胞分化,后者在分化过程中释放的TNF-α可能与ECV304细胞PCA活性升高有关;TanⅡA又能抑制TanⅡA-NB4-CM增强ECV304细胞PCA的作用。
Objective: To study the effect of Tan Ⅱ A on the procoagulant activity (PCA) induced by NB4 cells in acute promyelocytic leukemia (APL) cell line, and to explore its mechanism. Methods: (1) NB4 cells were treated with 1.0 μg / mL Tan Ⅱ A, 0.3 μg / mL ATRA, 0.01% DMSO and PRMI1640 for 24,48 and 72 h respectively. The supernatant was used as conditioned medium (hNB4-CM). The ECs were incubated with ECV304 cells at 37 ° C for 0, 4, 8 and 12 h, respectively. ECV304 cell lysate was prepared by repeated freeze-thaw cycles and the PCA was determined by one-stage coagulation assay. The TNF- α. (2) ECV304 cells were incubated with 1.0 μg / mL TanⅡA and TanⅡA 72h-NB4-CM at 37 ℃ for 6, 12, 24 and 48 hours, respectively. ATRA and DMSO were used as positive and negative controls, respectively. ECV304 cells Lysis solution of PCA. Results: (1) 1.0μg / mL TanⅡA could induce the NB4 cell differentiation. The effect of NB4 cells in medium increased the expression of PCA in ECV304 cells, which peaked at 4h after incubation. The PCA of ECV304 cells decreased gradually. There was no significant difference (P> 0.05) with 0.3μg / mL ATRA. (2) 1.0μg / mL TanⅡA inhibited the proliferation of PCA in ECV304 cells induced by TanⅡA-72h-NB4-CM, and its intensity increased with the increase of time. Compared with 1.0μmol / L ATRA, P> 0.05. (3) The concentration of TNF-α in medium of NBⅡcells induced by TanⅡA increased with the increase of time within 7h, which showed no difference with 0.3μg / mL ATRA (P> 0.05). CONCLUSION: TanⅡA can induce the differentiation of NB4 cells. The TNF-α released by the latter may be related to the increase of PCA activity in ECV304 cells. TanⅡA can inhibit the effect of TanⅡA-NB4-CM on enhancing the PCA of ECV304 cells.