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目的观察蓬子菜总黄酮(FGVL)诱导急性早幼粒细胞白血病细胞株(NB4)细胞凋亡作用,并探讨其作用机制。方法 FGVL(50、100、200μg/mL)作用于体外培养的NB4细胞,Hoechst染色观察细胞形态学变化;流式细胞术检测细胞周期,Annexin V-FITC/PI双标记法检测细胞凋亡状况;RT-PCR检测NB4凋亡相关基因Mel-18、Sall4、Bmi-1 m RNA表达。结果细胞形态学观察显示,NB4细胞经FGVL处理48 h后,与对照组比较,FGVL各剂量组NB4细胞凋亡比例均明显升高(P<0.05),呈现细胞核固缩、核碎裂及凋亡小体等典型的细胞凋亡特征;与对照组(2.3%)比较,50、100、200μg/mL FGVL组NB4细胞凋亡率(分别为9.3%、13%、20.4%)明显升高(P<0.05);细胞周期检测结果显示,与对照组比较,50、100、200μg/mL FGVL组NB4细胞G0/G1期细胞比例(分别为43.92%、37.26%、29.15%)减少,S期细胞比例(54.16%、62.23%、70.46%)增多;与对照组比较,100、200μg/mL FGVL组NB4细胞M el-18 m RNA表达水平升高,Sall4、Bmi-1 m RNA表达水平明显降低。结论 FGVL可阻滞NB4细胞于S期,抑制细胞增殖,通过调节M el-18、Sall4、Bmi-1等凋亡相关基因表达诱导细胞凋亡。
Objective To observe the apoptosis of acute promyelocytic leukemia cell line (NB4) induced by total flavonoids of Phragmites australis (FGVL) and to explore its mechanism. Methods FGVL (50, 100, 200μg / mL) was used to in vitro cultured NB4 cells. Hoechst staining was used to observe the morphological changes of cells. Flow cytometry was used to detect cell cycle. Annexin V-FITC / PI double- RT-PCR was used to detect the expression of Mel-18, Sall4 and Bmi-1 mRNA in NB4 cells. Results The results of morphological observation showed that compared with the control group, the apoptosis rate of NB4 cells in each dose group of FGVL increased significantly (P <0.05) after treated with FGVL for 48 h, showing nuclear pyknosis, nuclear fragmentation and apoptosis (3. 9%, 13%, 20.4%, respectively) in 50, 100, 200μg / mL FGVL group were significantly higher than those in control group (2.3% P <0.05). The results of cell cycle assay showed that compared with the control group, the percentage of cells in G0 / G1 phase of NB4 cells decreased (43.92%, 37.26% and 29.15%, respectively) in 50,100,200μg / mL FGVL group, (54.16%, 62.23%, 70.46%). Compared with the control group, the expression of M el-18 m RNA in NB4 cells in 100,200 μg / mL FGVL group was significantly increased and the expression of Sall4 and Bmi-1 m RNA was significantly decreased. Conclusion FGVL can block NB4 cells in S phase and inhibit cell proliferation, and induce apoptosis by regulating the expression of apoptosis-related genes such as M el-18, Sall4 and Bmi-1.