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目的探讨苦参碱(Mat)诱导急性早幼粒白血病细胞株NB4细胞凋亡作用及机制。方法不同剂量M at(0.25、0.50、1.00 mg/m L)作用于NB4细胞(24、48、72 h),噻唑蓝法(M TT)检测细胞增殖抑制率;Hoechst33258染色观测细胞形态学变化;流式细胞术(FCM)检测细胞周期,Annexin V-FITC/PI双标记法测定细胞凋亡率;分光光度法检测NB4细胞内caspase-3和caspase-8活性。结果与对照组比较,Mat可明显抑制NB4细胞增殖,诱导细胞凋亡,呈时间和剂量效应(P<0.05);细胞形态学观察可见明显的细胞凋亡特征;与对照组比较,0.50、1.00 mg/m L M at组G0/G1期细胞比例(分别为57.91%、83.00%)明显增加,S期比例(分别为40.95%、16.10%)减少;与对照组比较,0.50、1.00 mg/m L Mat组NB4细胞内caspase-3酶活性[分别为(77.41±4.01)、(111.78±4.05)]、1.00 mg/m L M at组NB4细胞内caspase-8酶活性(44.98±7.63)明显增强(P<0.05)。结论 M at可抑制NB4细胞增殖并诱导细胞凋亡,其机制可能与G_0/G_1期细胞阻滞、抑制DNA合成并激活细胞内caspase-3、caspase-8凋亡途径有关。
Objective To investigate the effect of matrine on the apoptosis of acute promyelocytic leukemia NB4 cells and its mechanism. Methods The cells were treated with various doses of M at (0.25, 0.50, 1.00 mg / m L) for 24,48 and 72 h. MTT assay was used to detect the cell proliferation inhibition rate. Morphological changes were observed by Hoechst33258 staining. The cell cycle was detected by flow cytometry (FCM). The apoptosis rate was determined by Annexin V-FITC / PI double labeling method. The activity of caspase-3 and caspase-8 in NB4 cells was detected by spectrophotometry. Results Compared with the control group, Mat significantly inhibited the proliferation of NB4 cells and induced the apoptosis in time and dose-dependent manner (P <0.05). The morphological characteristics of cells showed obvious apoptosis characteristics. Compared with the control group, 0.50, 1.00 Compared with the control group, the proportion of cells in the G0 / G1 phase (57.91% and 83.00%, respectively) was significantly increased in the mg / m LM at group and decreased in the S phase (40.95% and 16.10% The activity of caspase-3 in NB4 cells in the Mat group (77.41 ± 4.01, (111.78 ± 4.05, respectively)] was significantly increased (P <0.05) <0.05). Conclusions M at can inhibit the proliferation and induce the apoptosis of NB4 cells. The mechanism may be related to the cell arrest of G 0 / G 1, the inhibition of DNA synthesis and the activation of caspase-3 and caspase-8.