槲皮素对白血病细胞杀伤作用的研究  

The study of killing effect of quercetin on leukemia cells

作　　者：黄琰[1] 吴隼[1] 张媛[1] 字友梅[1] 杨满[1] 郭燕[1] 张灵秀[1] 王莉华[1] 

机构地区：[1]新乡医学院第一附属医院,卫辉453100

出　　处：《中药药理与临床》2015年第5期61-65,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河南省教育厅基金资助项目(编号:2006320039)

摘　　要：目的：研究槲皮素对白血病细胞的杀伤作用以及诱导的凋亡研究。方法：CCK-8试剂盒检测不同浓度的槲皮素（0～100μM）对白血病K562和HL-60细胞活性的影响;Hoechst33342染色技术观察40u M槲皮素处理细胞24和48小时后细胞核形态的变化;流式细胞仪检测细胞凋亡情况;caspase8、9、3（半胱天冬酶）特异性抑制剂预处理K562细胞后,检测药物对细胞活性的影响;40u M槲皮素处理细胞24和48小时后,Western blot检测细胞中caspase家族蛋白8、9、3的表达情况。结果：0～100u M的槲皮素能够引起K562和HL-60细胞剂量依赖型的活性下降,40u M的槲皮素处理细胞24和48小时后,K562细胞核明显发生皱缩,引起凋亡,随着处理时间增长,凋亡率增加,处理24和48小时后细胞的凋亡率分别为（45.8±2.06）%和（83.6±1.89）%。caspase 9、3抑制剂预处理K562细胞后,槲皮素能够引起细胞活性的增加,而caspase 8抑制剂预处理后,槲皮素没有引起细胞活性的上升。Western blot检测结果表明40μM的槲皮素处理细胞后能够上调caspase 9、3蛋白的表达,而没有引起caspase 8蛋白表达量的上升,随着处理时间的增长,caspase 9、3的表达量更高。结论：槲皮素对白血病细胞具有很强的杀伤作用,并诱导细胞caspase 9、3蛋白参与的内源性通路介导的凋亡。Objective： To investigate the killing effect of quercetin on leukemia cells and the cell apoptosis induced by quercetin. Methods： CCK-8 assays was used to detect the viability of leukemia cell K562 and HL-60 after treating with different concentration of quercetin（ 0 ～ 100 u M）; Hoechst33342 was used to stain the cell to observe the nucleus change; flow cytometry was further used to test the K562 cell apoptosis and its apoptotic ratio; after pretreatment with caspase8,9,3 inhibitors,the cell viability was detected; at last,western blot was used to detect the caspase8,9,3 protein expression of K562 cell after treating with 40 u M quercetin for 24 and 48 h. Results： 0 ～ 100 u M of quercetin could induce the dose-dependent viability decrement of K562 and HL-60 cells（ p 0. 05）. Compared with control group,after treating with40 u M of quercetin for 24 and 48 h,K562 cells showed significant nucleus pyknosis and further apoptosis,which the apoptotic ratio was（ 45.8 ± 2. 06） % and（ 83. 6 ± 1. 89） %（ p 0. 01） respectively. After pretreatment with caspase 9,3 inhibitors,quercetin could enhance the cell viability compared with single quercetin treating group（ p 0. 01）,but caspase 8 didn ＇t had similar effect. At last,western blot results showed that quercetin could up-regulate the caspase 9 and 3 protein expression but not caspase 8 protein. Conclusions： Caspase protein family members 9,3 joined the quercetin induced leukemia cell apoptosis. Quercetin had strong killing effect on leukemia cells and induced endogenous pathways mediated apoptosis.

关 键 词：槲皮素 白血病细胞 caspase蛋白家族 凋亡 

分 类 号：R285[医药卫生—中药学]