富硒麦芽粉水提取物通过细胞衰老和凋亡抑制人白血病K562细胞的增殖

The proliferation-inhibited action of water extracts from selenium-enriched malt powder in K562 cells through cellular senescence and apoptosis

目的 研究富硒麦芽粉水提取物能否抑制K562细胞增殖的作用.方法 Coulter细胞计数法检测细胞增殖的抑制作用,流式细胞仪检测细胞周期分布以及细胞内活性氧自由基的水平,DNA特异染料Hoechst 33342染色检测染色质凝集,免疫印迹法检测凋亡相关蛋白的表达,衰老相关的β-半乳糖苷酶染色检测细胞衰老.结果 富硒麦芽粉水提取物能抑制K562细胞的增殖,IC50值为0.5 mg/mL.进一步研究发现：富硒麦芽粉水提取物使细胞阻滞在G2/M期、增加细胞内活性氧自由基水平,染色质发生凝集,引起细胞凋亡标志蛋白的切割.此外,4 mg/mL的水提取物能引起60％的细胞发生衰老.结论 富硒麦芽粉水提取物具有明显的抑制K562细胞增殖的作用,其机制与诱导细胞衰老和凋亡有关.

Objective To investigate whether the water extracts from selenium-enriched malt powder （WSM） can inhibit the proliferation of K562 cells. Methods Coulter cell counting method was used to detect the inhibitory effect on cell proliferation. Cell cycle distribution and accumulation of intracellular reactive oxygen species （ROS） were analyzed with flow cytometry. Chromatin condensation was detected by a fluorescent microscope after Hoechst 33342 staining. Protein expression was detected by immune-blotting. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Results WSM exhibited the inhibition action on K562 cells with 0.5 mg/mL of the IC50 value. Furthermore, the arrest at G2/M phase, an increased ROS level, chromatin condensation and cleavage of the marker protein of apoptosis were detected after exposure to WSM. In addition, about 60 % of the cells were occurred cellular senescence after the treat- ment with 4 mg/mL of WSM. Conclusion WSM can inhibit the proliferation of K562 cells, and its mechanism is related to the induction of cellular senescence and apoptosis.